{"id":6758,"date":"2024-09-24T23:24:09","date_gmt":"2024-09-24T15:24:09","guid":{"rendered":"https:\/\/fanyiming.life\/?p=6758"},"modified":"2025-12-21T01:04:20","modified_gmt":"2025-12-20T17:04:20","slug":"chip-seq-protocol","status":"publish","type":"post","link":"https:\/\/fanyiming.life\/?p=6758","title":{"rendered":"ChIP-seq Protocol"},"content":{"rendered":"\t<div id=\"Experiment-Process\">\r\n    <!-- \u6dfb\u52a0\u6309\u94ae -->\r\n        <div class=\"day-buttons\" style=\"padding-left: 10px;\">\r\n            <button class=\"ChIP-button\" onclick=\"showDay('day1')\">Day 1<\/button>\r\n            <button class=\"ChIP-button\" onclick=\"showDay('day2')\">Day 2<\/button>\r\n            <button class=\"ChIP-button\" onclick=\"showDay('day3')\">Day 3<\/button>\r\n            <button class=\"ChIP-button\" onclick=\"showDay('day4')\">Day 4<\/button>\r\n            <button class=\"ChIP-button\" onclick=\"showDay('reagent')\">BufferRecipes<\/button>\r\n            <button class=\"ChIP-button\" onclick=\"showDay('reagentCalc')\">ReagentCalc<\/button>\r\n        <\/div>\r\n        <div id=\"day1\" class = \"ChIP-day\">\r\n            <h3 >Day1<\/h3>\r\n            <!-- Step 1 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 1: \u7ec6\u80de\u6536\u96c6\u548c\u4ea4\u8054<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u5728\u4e24\u4e2a6\u5b54\u677f\u7684\u5b54\u4e2d\u57f9\u517b\u7ec6\u80de\u81f3\u7ea680%\u7684\u6c47\u5408\u5ea6\uff08~<strong style=\"color: blue;\">5million<\/strong>\u7ec6\u80de\uff09\uff0c\u7528<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>1x PBS\u6e05\u6d17\u4e00\u6b21\uff0c\u5728<strong style=\"color:orange;\">37\u2103<\/strong>\u4e0b\u7528\u80f0\u86cb\u767d\u9176\u6d88\u5316<strong style=\"color: #CC1276;\">3\u5206\u949f<\/strong>\u540e\uff0c\u6536\u96c6\u81f32 mL\u4f4e\u5438\u9644\u7ba1\u4e2d\uff0c\u52a0\u5165<strong style=\"color: green;\">1 mL<\/strong>\u57f9\u517b\u57fa\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5bf9\u6536\u96c6\u5230\u7684\u7ec6\u80de\u60ac\u6db2\u8fdb\u884c\u7ec6\u80de\u8ba1\u6570\u3002\u5bf9\u4e8eNCCIT\u7ec6\u80de\u7cfb\uff0c\u6839\u636e\u7ec6\u80de\u8ba1\u6570\u7ed3\u679c\u53d6\u7528<strong style=\"color: blue;\">3 million<\/strong>\u7ec6\u80de\u5bf9\u5e94\u7684\u4f53\u79ef\u7684\u7ec6\u80de\u60ac\u6db2\uff0c<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u79bb\u5fc3\u6536\u96c6\u3002<strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\uff0c<strong style=\"color: #CC1276;\">200xg<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u79bb\u5fc3\u65f6\u914d\u7f6e<strong style=\"color: blue;\">3% PFA in PBS<\/strong>\uff08\u6bcf<strong style=\"color: green;\">1.5mL<\/strong> PBS\u52a0<strong style=\"color: green;\">300\u03bcL<\/strong> 16% PFA\uff0c\u4ee5\u8fbe\u5230\u6700\u7ec8<strong style=\"color: blue;\">3% PFA in PBS<\/strong>\u7684\u6d53\u5ea6\uff09\u3002\u79bb\u5fc3\u7ed3\u675f\u540e\uff0c\u5f03\u4e0a\u6e05\uff0c\u7528<strong style=\"color: blue;\">1.8mL 3% PFA in PBS<\/strong>\u91cd\u60ac\uff08\u5148\u52a0<strong style=\"color: green;\">900\u03bcL<\/strong>\u5145\u5206\u91cd\u60ac\u540e\u518d\u52a0\u53e6\u5916\u7684<strong style=\"color: green;\">900\u03bcL<\/strong>\uff09\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        <strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\uff0c\u5782\u76f4\u65cb\u8f6c<strong style=\"color: #CC1276;\">10\u5206\u949f<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u8fc7\u7a0b\u4e2d\u914d\u7f6e\u4e0b\u4e00\u6b65\u9700\u8981\u7684\u53cd\u5e94\u7ec8\u6b62\u6db2\uff08<strong style=\"color: purple;\">0.94g<\/strong> Glycine + <strong style=\"color: green;\">5mL<\/strong> H<sub>2<\/sub>O\uff09\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n                <p><strong style=\"color: #CC1276;\">\u6ce8\u610f\uff1a<\/strong><strong>1.5mL PBS + 300\u03bcL 16% PFA \u7684\u5b9e\u9645\u7ec8\u6d53\u5ea6\u4e3a 2.67%\uff0c\u4f46\u5b9e\u9a8c\u5ba4\u4e60\u60ef\u79f0\u547c\u4e3a 3% PFA\u3002<\/strong><\/p>\r\n            <\/div>\r\n\r\n            <!-- Step 2 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 2: \u7ec8\u6b62\u4ea4\u8054<\/h4>\r\n                <ol>\r\n                    <li>\u52a0\u5165<strong style=\"color: green;\">180uL<\/strong> 2.5M\u7518\u6c28\u9178\uff0c\u5feb\u901f\u4e0a\u4e0b\u5439\u62533\u6b21\uff0c\u8f7b\u8f7b\u98a0\u5012\u51e0\u6b21\u6df7\u5408\u540e\uff0c<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u5782\u76f4\u65cb\u8f6c<strong style=\"color: #CC1276;\">10\u5206\u949f<\/strong>\u3002<\/li>\r\n                <\/ol>            <p><strong style=\"color: #CC1276;\">\u63d0\u793a\uff1a<\/strong> <\/p>\r\n                <ul>\r\n                    <li>\r\n                        <strong style=\"color: blue;\">2.5M<\/strong> \u7518\u6c28\u9178\u7528\u4e8e <strong style=\"color: blue;\">3%<\/strong> PFA \u4ea4\u8054\uff0c\u914d\u65b9\uff1a<strong style=\"color: purple;\">0.94g<\/strong> \u7518\u6c28\u9178 + <strong style=\"color: green;\">5mL<\/strong> H<sub>2<\/sub>O\uff1b\r\n                    <\/li>\r\n                    <li>\r\n                        <strong style=\"color: blue;\">1.25M<\/strong> \u7518\u6c28\u9178\u7528\u4e8e <strong style=\"color: blue;\">1%<\/strong> PFA \u4ea4\u8054\uff0c\u914d\u65b9\uff1a<strong style=\"color: purple;\">0.47g<\/strong> \u7518\u6c28\u9178 + <strong style=\"color: green;\">5mL<\/strong> H<sub>2<\/sub>O\u3002\r\n                    <\/li>\r\n                <\/ul>        \r\n            <\/div>\r\n\r\n            <!-- Step 3 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 3: PBS\u6e05\u6d17<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u79bb\u5fc3<strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\uff0c\r\n                        <strong style=\"color: #CC1276;\"><strong style=\"color: blue;\">4\u2103<\/strong><\/strong>\uff0c\r\n                        <strong style=\"color: #CC1276;\">8000xg<\/strong>\uff0c\r\n                        \u5f03\u4e0a\u6e05\uff0c\u52a0<strong style=\"color: green;\">1 mL<\/strong>\u51b7\u76841x PBS\uff0cVortex\u6df7\u5300\r\n                        <i class = \"ChIP-note\"><br>\u8fd9\u91cc\u53ef\u4ee5\u53d1\u73b0\u7ec6\u80de\u6c89\u6dc0\u6b7b\u6b7b\u9ecf\u5728\u79bb\u5fc3\u7ba1\u5e95\u90e8\uff0c\u6bd4\u8f83\u96beVortex\u8d77\u6765\uff0c\u53ef\u4ee5\u7528\u624b\u6307\u5934\u5f39\u4e00\u5f39\u5e2e\u52a9\u6df7\u5300\u3002<\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        \u518d\u6b21\u79bb\u5fc3<strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\uff0c\r\n                        <strong style=\"color: #CC1276;\"><strong style=\"color: blue;\">4\u2103<\/strong><\/strong>\uff0c\r\n                        <strong style=\"color: #CC1276;\">8000xg<\/strong>\uff0c\r\n                        \u5f03\u4e0a\u6e05\uff0c\u91cd\u60ac\u4e8e<strong style=\"color: green;\">1 mL<\/strong>\u51b7\u76841x PBS\uff0cVortex\u6df7\u5300\r\n                        <i class = \"ChIP-note\"><br>\u8fd9\u91cc\u6bd4\u4e0a\u4e00\u6b65Vortex\u5c31\u8f7b\u677e\u591a\u4e86\u3002<\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        \u518d\u6b21\u79bb\u5fc3<strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\uff0c\r\n                        <strong style=\"color: #CC1276;\"><strong style=\"color: blue;\">4\u2103<\/strong><\/strong>\uff0c\r\n                        <strong style=\"color: #CC1276;\">8000xg<\/strong>\uff0c\r\n                        \u5f03\u4e0a\u6e05\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 4 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 4: \u88c2\u89e3\u7f13\u51b2\u6db2LB1\u548cLB2\u5904\u7406<\/h4>\r\n                <ol>\r\n                    <li>\u91cd\u60ac\u7ec6\u80de\u6c89\u6dc0\u4e8e<strong style=\"color: green;\">2 mL<\/strong>\u51b7\u7684LB1\u7f13\u51b2\u6db2\u4e2d\uff0c\u8f7b\u8f7b\u4e0a\u4e0b\u5439\u6253\u540e\uff0c<strong style=\"color: #CC1276;\"><strong style=\"color:blue;\">4\u2103<\/strong><\/strong>\u4e0b\u5782\u76f4\u65cb\u8f6c<strong style=\"color: #CC1276;\">10\u5206\u949f<\/strong>\u3002<\/li>\r\n                    <li>\r\n                        \u79bb\u5fc3<strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\uff0c<strong style=\"color: #CC1276;\">8000xg<\/strong>\uff0c<strong style=\"color: #CC1276;\"><strong style=\"color:blue;\">4\u2103<\/strong><\/strong>\uff0c\u5f03\u4e0a\u6e05\u540e\u7528\u5c0f\u67aa\u5934\u5438\u53bb\u6b8b\u4f59\u6db2\u4f53\r\n                        <i class = \"ChIP-note\"><br>\u8fd9\u91cc\u6c89\u6dc0\u7c98\u9644\u4e0d\u5f3a\uff0c\u9700\u8981\u5f88\u5c0f\u5fc3\u5730\u5438\u5f03\u4e0a\u6e05<\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        \u91cd\u60ac\u7ec6\u80de\u6c89\u6dc0\u4e8e<strong style=\"color: green;\">2 mL<\/strong><strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>LB2\u7f13\u51b2\u6db2\uff0c\u8f7b\u8f7b\u4e0a\u4e0b\u5439\u6253\uff0c<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u5782\u76f4\u65cb\u8f6c<strong style=\"color:#CC1276;\">10\u5206\u949f<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u79bb\u5fc3<strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\uff0c<strong style=\"color: #CC1276;\">8000xg<\/strong>\uff0c<strong style=\"color: #CC1276;\"><strong style=\"color:blue;\">4\u2103<\/strong><\/strong>\uff0c\u5f03\u4e0a\u6e05\u540e\u7528\u5c0f\u67aa\u5934\u5438\u53bb\u6b8b\u4f59\u6db2\u4f53\u3002\r\n                        <i class = \"ChIP-note\"><br>\u8fd9\u91cc\u6c89\u6dc0\u7c98\u9644\u4ecd\u7136\u4e0d\u5f3a\uff0c\u9700\u8981\u5f88\u5c0f\u5fc3\u5730\u5438\u5f03\u4e0a\u6e05<\/i>\r\n                    <\/li>\r\n                <\/ol>    <\/div>\r\n            <!-- Step 5 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 5: \u8d85\u58f0\u526a\u5207DNA<\/h4>\r\n                \r\n                <ol>\r\n                    <li>\r\n                        \u51c6\u5907\u8d85\u58f0\u4e13\u7528\u7ba1\u3002\u91cd\u60ac\u7ec6\u80de\u6c89\u6dc0\u4e8e<strong style=\"color: green;\">301uL<\/strong>\u51b7\u7684Shearing Buffer 1 (with 0.3% SDS)\u4e2d\uff0c\u8f7b\u8f7b\u4e0a\u4e0b\u5439\u6253\u540e\u8f6c\u79fb\u5230\u8d85\u58f0\u4e13\u7528\u7ba1\uff0c\u7f6e\u4e8e\u51b0\u4e0a\r\n                        <i class = \"ChIP-note\"><br>\u6709SDS\u5bb9\u6613\u4ea7\u751f\u6c14\u6ce1\uff0c\u53ef\u4ee5\u75281mL\u79fb\u6db2\u67aa\u60ac\u7a7a\u6ef4\u843d\uff0c\u540e\u7528200\u03bcL\u79fb\u6db2\u67aa\u7f13\u6162\u6376\u6253\u6df7\u5300\u3001\u8f6c\u79fb<\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        \u8d85\u58f0\u3002<strong style=\"color: blue;\">\u8d85\u58f0\u53c2\u6570\uff1a<\/strong> \u632f\u5e45 <strong style=\"color: blue;\">85%<\/strong>\uff0c<strong style=\"color: blue;\">20\u79d2\u5f00\/40\u79d2\u5173<\/strong>\uff0c\u5904\u7406<strong style=\"color: #CC1276;\">22\u5206\u949f<\/strong>\r\n                        <i class = \"ChIP-note\">\r\n                            <br>\u5bf9\u4e8eNCCIT\u7ec6\u80de\u8bbe\u7f6e22min\uff0c\u5b9e\u9645\u52a0\u4e0a\u6682\u505c\u7684\u65f6\u95f4\u4e00\u5171\u662f66min\uff1b\u5bf9\u4e8eK562\u9700\u8981\u8bbe\u7f6e25min\r\n                        <\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        \u6bcf\u4e2a\u8d85\u58f0\u540e\u7684\u6837\u54c1\u8f6c\u79fb\u81f31.5 mL\u9884\u51b7\u7ba1\uff0c\u52a0\u5165<strong style=\"color: green;\">602uL<\/strong>Shearing Buffer 2\uff08no SDS\uff09\uff0c\u79bb\u5fc3<strong style=\"color: #CC1276;\">10\u5206\u949f<\/strong>\uff0c<strong style=\"color: #CC1276;\">16000xg<\/strong>\uff0c<strong style=\"color: #CC1276;\"><strong style=\"color:blue;\">4\u2103<\/strong><\/strong>\r\n                        <i class = \"ChIP-note\">\r\n                            <br>\u8fd9\u91cc\u8d85\u58f0\u5b8c\u7ba1\u58c1\u4f1a\u6709\u5927\u91cf\u5c0f\u6c34\u73e0\uff0c\u9700\u8981\u5bf9\u8d85\u58f0\u7ba1\u77ac\u65f6\u79bb\u5fc3\u4e00\u4e0b\u518d\u8f6c\u79fb\r\n                        <\/i>\r\n                    <\/li>\r\n                <\/ol>        <\/div>\r\n\r\n            <!-- Step 6 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 6: \u5b75\u80b2\u6297\u4f53<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u8f6c\u79fb\u4e0a\u6e05\u6db2\uff08\u7ea6<strong style=\"color: green;\">900uL<\/strong>\uff09\u81f3\u65b0\u76841.5 mL\u9884\u51b7\u7ba1\u4e2d\uff0c\u52a0\u5165<strong style=\"color: green;\">100uL<\/strong> 10% Triton X-100\uff0c\u6df7\u5300\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u53d6<strong style=\"color: green;\">5\u03bcL<\/strong>\u81f3\u65b0\u76841.5mL\u4f4e\u5438\u9644\u79bb\u5fc3\u7ba1\u4f5c\u4e3a<strong>\"input\"<\/strong>\uff0c-20\u2103\u5b58\u653e\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5269\u4f59\u6837\u54c1\u5206\u6210\u4e24\u4e2a<strong>replicate<\/strong>\uff0c\u6bcf\u4efd<strong style=\"color: green;\">450\u03bcL<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u6bcf\u4e2a<strong>replicate<\/strong>\u4e2d\u52a0\u5165\u7ea6<strong style=\"color: purple;\">5\u03bcg<\/strong> \u6297\u4f53\uff0c\u6df7\u5408\u540e\u5728<strong style=\"color:blue;\">4\u2103<\/strong>\u4e0b\u5782\u76f4\u65cb\u8f6c<strong style=\"color: #CC1276;\">12-16\u5c0f\u65f6<\/strong>\u3002\r\n                    <\/li>\r\n                <\/ol>        \r\n                <p><strong style=\"color: #CC1276;\">\u6ce8\u610f\uff1a<\/strong> <strong style=\"color: purple;\">5\u03bcg<\/strong> \u6297\u4f53\u662f\u53c2\u8003\u503c\uff0c\u4f9d\u4e0d\u540c\u6297\u4f53\u548c\u86cb\u767d\u800c\u53d8\u5316\u3002<\/p> \r\n            <\/div>\r\n\r\n        <\/div>\r\n        <div id=\"day2\" class = \"ChIP-day\">\r\n            <h3 >Day2<\/h3>\r\n            <!-- Step 7 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 7: \u51c6\u5907Dynabeads\u5e76\u6d17\u6da4<\/h4>\r\n            <ol>\r\n                <li>\r\n                    \u786e\u5b9a\u5e94\u4f7f\u7528\u54ea\u79cd\u78c1\u6027Dynabeads\uff08Protein A\u6216Protein G\uff09\u4e0e\u5404\u7c7b\u6297\u4f53\u7ed3\u5408 \r\n                    [\u53c2\u9605Dynabeads\u624b\u518c\u4e2d\u7684\u6297\u4f53\u7c7b\u578b\u4eb2\u548c\u529b]\u3002\r\n                <\/li>\r\n                <li>\r\n                    \u6bcf\u4e2aChIP\u53cd\u5e94\u7b49\u5206 <strong style=\"color: green;\">50 \u00b5L<\/strong> Dynabead\u60ac\u6db2\uff08\u76f8\u5f53\u4e8e <strong style=\"color: green;\">25 \u00b5L<\/strong> \u78c1\u73e0\uff09\u81f3\u7f6e\u4e8e\u51b0\u4e0a\u76841.5mL\u79bb\u5fc3\u7ba1\u4e2d\uff08\u8fd9\u91cc\u6307\u7684\u662f\u6bcf\u4e2areplicate\uff09\u3002\r\n                <\/li>\r\n                <li>\r\n                    \u5728\u51b0\u4e0a\u7528<strong style=\"color: green;\">1 mL<\/strong>\u51b7\u7684Block Solution\u5c06Dynabeads\u6d17\u6da4<strong>3\u6b21<\/strong>\uff1a\r\n                    <ol class=\"sub-list\">\r\n                        <li>\u52a0\u5165 <strong style=\"color: green;\">1 mL<\/strong> \u51b7\u7684Block Solution\uff0c\u901a\u8fc7\u98a0\u5012\u6216\u8f7b\u5f39\u6df7\u5300<\/li>\r\n                        <li>\u653e\u5165\u78c1\u529b\u67b6\uff0c\u7b49\u5f85\u78c1\u73e0\u9644\u7740\u5728\u4fa7\u58c1<\/li>\r\n                        <li>\u5438\u8d70\u6db2\u4f53\uff0c\u91cd\u590d\u6b64\u8fc7\u7a0b<\/li>\r\n                        <li>\u6700\u540e\u4e00\u6b21\u6d17\u6da4\u65f6\uff0c\u7528\u5c0f\u67aa\u5934\u79fb\u9664\u6240\u6709\u6db2\u4f53<\/li>\r\n                    <\/ol>\r\n                <\/li>\r\n            <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 8 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 8: \u6297\u4f53\u7ed3\u5408<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u5c06\u6240\u6709\u7ea6 <strong style=\"color: green;\">0.45 mL<\/strong> \u6297\u4f53\u7ed3\u5408\u7684\u67d3\u8272\u8d28\uff08\u5c31\u662f\u4e0a\u4e00\u6b654\u2103\u5b75\u80b2\u7684\u6837\u54c1\uff09\u52a0\u5165\u5df2\u6d17\u6da4\u7684\u78c1\u73e0\u4e2d\uff0c\u98a0\u5012\u6df7\u5408\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728 <strong style=\"color:blue;\">4\u2103<\/strong> \u4e0b\u5782\u76f4\u65cb\u8f6c\u81f3\u5c11 <strong style=\"color: #CC1276;\">2-4\u5c0f\u65f6<\/strong>\uff0c\u4f7f\u6297\u4f53\u7ed3\u5408\u5230\u78c1\u73e0\u4e0a\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 9 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 9: \u6d17\u6da4\u7ed3\u5408\u7684\u78c1\u73e0<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u51b0\u4e0a\u51b7\u5374RIPA Wash Buffer\uff0c\u5e76\u5c06\u6c34\u6d74\u8bbe\u7f6e\u4e3a <strong style=\"color:rgb(255, 105, 59);\">65\u2103<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u7528 <strong style=\"color: green;\">1 mL<\/strong> \u51b7\u7684RIPA\u81f3\u5c11\u6d17\u6da4\u7ed3\u5408\u7684\u78c1\u73e0<strong>3\u6b21<\/strong>\uff1a\r\n                        <ol class = \"sub-list\">\r\n                            <li>\u52a0\u5165 <strong style=\"color: green;\">1 mL<\/strong> \u51b7\u7684RIPA\uff0c\u98a0\u5012\u6216\u8f7b\u5f39\u6df7\u5300<\/li>\r\n                            <li>\u653e\u5165\u78c1\u529b\u67b6\uff0c\u7b49\u5f85\u78c1\u73e0\u9644\u7740\u5728\u4fa7\u58c1<\/li>\r\n                            <li>\u5438\u8d70\u6db2\u4f53\uff0c\u91cd\u590d\u6b64\u8fc7\u7a0b<\/li>\r\n                            <li>\u6700\u540e\u4e00\u6b21\u6d17\u6da4\u65f6\uff0c\u7528\u5c0f\u67aa\u5934\u79fb\u9664\u6240\u6709\u6db2\u4f53<i class = \"ChIP-note\"><br>\u56e0\u4e3a\u6709NP-40\uff0cRIPA Wash Buffer\u6709\u4e00\u70b9\u70b9\u7c98\uff0c\u6240\u4ee5\u7528\u5927\u67aa\u5438\u5b8c\u540e\u4f1a\u6709\u4e00\u4e9b\u7ba1\u58c1\u4e0a\u7684\u6db2\u4f53\u6162\u60a0\u60a0\u518d\u843d\u5230\u7ba1\u5e95\u90e8\uff0c\u8fd9\u65f6\u518d\u7528\u5c0f\u67aa\u5934\u5438\u5e72\uff08\u4e0d\u8981\u5927\u67aa\u5934\u4e0b\u9762\u6233\u4e00\u4e2a\u5c0f\u67aa\u5934\u53ea\u5438\u4e00\u6b21\uff0c\u90a3\u6837\u8fd8\u662f\u4f1a\u6709\u6b8b\u7559\uff09\u3002<\/i><\/li>\r\n                        <\/ol>\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728\u51b0\u4e0a\u7528 <strong style=\"color: green;\">1 mL<\/strong> \u542b <strong style=\"color: blue;\">50 mM<\/strong> NaCl \u7684TE Buffer\u6d17\u6da4\u78c1\u73e0<strong>1\u6b21<\/strong>\uff0c\u653e\u5165\u78c1\u529b\u67b6\uff0c\u7b49\u5f85\u78c1\u73e0\u9644\u7740\u5728\u4fa7\u58c1\u540e\u5438\u8d70\u6db2\u4f53\u3002\r\n                        <i class = \"ChIP-note\"><br>\u75285M NaCl\u548cTE Buffer\u73b0\u914d\u73b0\u7528\u3002\u7528TE+NaCl\u6d17\u7684\u65f6\u5019\uff0c\u660e\u663e\u611f\u89c9\u4e0d\u50cf\u524d\u9762RIPA Wash Buffe\u90a3\u6837\u98a0\u5012\u51e0\u6b21\u5c31\u80fd\u6df7\u5300\u4e86\uff0c\u4e00\u4e9bbeads\u6bd4\u8f83\u7262\u56fa\u5730\u7c98\u5728\u5e95\u90e8\u3002<\/i>\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 10 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 10: \u6d17\u8131\u67d3\u8272\u8d28<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u5728 <strong style=\"color:blue;\">4\u2103<\/strong> \u4e0b\u4ee5 <strong style=\"color: #CC1276;\">950 x g<\/strong> \u79bb\u5fc3 <strong style=\"color: #CC1276;\">3\u5206\u949f<\/strong>\uff0c\u7528\u5c0f\u67aa\u5934\u79fb\u9664\u6240\u6709\u5269\u4f59\u7684TE Buffer\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u52a0\u5165 <strong style=\"color: green;\">200 \u00b5L<\/strong> <strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u7684Elution Buffer\uff0c\u77ed\u6682vortex\u6df7\u5408\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728 <strong style=\"color:rgb(255, 105, 59);\">65\u2103<\/strong> \u7684\u6052\u6e29\u6df7\u5408\u5668\uff08Eppendorf Thermomixer C\uff09\u4e2d\u5b75\u80b2 <strong style=\"color: #CC1276;\">15\u5206\u949f<\/strong>\uff0c\u901f\u5ea6\u4e3a <strong style=\"color: #CC1276;\">1000 rpm<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u77ac\u65f6\u79bb\u5fc3 <strong style=\"color: #CC1276;\">10\u79d2<\/strong>\u9632\u6b62\u7ba1\u76d6\u4e0a\u6709\u6b8b\u4f59\u7684\u6837\u54c1\uff0c\u5c06\u78c1\u73e0\u653e\u5165\u78c1\u529b\u67b6\u4e2d\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u78c1\u73e0\u5b8c\u5168\u8d34\u58c1\u540e\uff0c\u5c06\u4e0a\u6e05\u6db2\uff08\u7ea6 <strong style=\"color: green;\">200 \u00b5L<\/strong>\uff09\u8f6c\u79fb\u5230\u65b0\u76841.5mL\u79bb\u5fc3\u7ba1\u4e2d\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 11 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 11: \u89e3\u4ea4\u8054<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u4ece <strong style=\"color:blue;\">-20\u2103<\/strong> \u5c06 <strong>\"input\"<\/strong> \u6837\u54c1\u89e3\u51bb\u81f3\u5ba4\u6e29\uff0c\u52a0\u5165Elution Buffer\u81f3 <strong style=\"color: green;\">200 \u00b5L<\/strong> \u4f53\u79ef\uff0c\u5e76\u901a\u8fc7\u77ed\u6682vortex\u6df7\u5408\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728 <strong style=\"color:rgb(255, 105, 59);\">65\u2103<\/strong> \u6052\u6e29\u6df7\u5408\u5668\uff08Eppendorf Thermomixer C\uff09\u4e2d\u5b75\u80b2ChIP\/input\u6837\u54c1\u8fc7\u591c\uff08 <strong style=\"color: #CC1276;\">12-16\u5c0f\u65f6<\/strong>\uff09\uff0c\u901f\u5ea6\u4e3a <strong style=\"color: #CC1276;\">300 rpm<\/strong>\uff0c\u4ee5\u89e3\u4ea4\u8054\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n        <\/div>\r\n        <div id=\"day3\" class = \"ChIP-day\">\r\n            <h3 >Day3<\/h3>\r\n            <!-- Step 12 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 12: \u7a00\u91caSDS\u548c\u6dfb\u52a0RNase A<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u5728<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u6bcf\u7ba1\u52a0\u5165 <strong style=\"color: green;\">1\u500d\u4f53\u79ef\uff08200 \u00b5L\uff09<\/strong> TE Buffer\uff08\u4ee5\u7a00\u91caSDS\uff09\uff0c\u5e76Vortex\u6df7\u5408\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u52a0\u5165 <strong style=\"color: green;\">8 \u00b5L<\/strong> \u6d53\u5ea6\u4e3a <strong style=\"color: green;\">10 mg\/mL<\/strong> \u7684RNase A\uff0c\u4f7f\u5176\u7ec8\u6d53\u5ea6\u4e3a <strong style=\"color: blue;\">0.2 mg\/mL<\/strong>\uff0c\u5e76Vortex\u6df7\u5408\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728 <strong style=\"color:orange;\">37\u2103<\/strong> \u7684\u6c34\u6d74\u6216\u6052\u6e29\u6df7\u5408\u5668\uff08Eppendorf Thermomixer C\uff09\u4e2d\u5b75\u80b2 <strong style=\"color: #CC1276;\">2\u5c0f\u65f6<\/strong>\uff0c\u901f\u5ea6\u4e3a <strong style=\"color: #CC1276;\">300 rpm<\/strong>\uff0c\u4ee5\u6d88\u5316RNA\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 13 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 13: \u6dfb\u52a0CaCl<sub>2<\/sub>\u548cProteinase K<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u52a0\u5165 <strong style=\"color: green;\">7 \u00b5L<\/strong> \u6d53\u5ea6\u4e3a <strong style=\"color: blue;\">300 mM<\/strong> \u7684CaCl<sub>2<\/sub>\uff0c\u4f7f\u5176\u7ec8\u6d53\u5ea6\u8fbe\u5230 <strong style=\"color: blue;\">5.25 mM<\/strong>\uff0c\u5e76Vortex\u6df7\u5408\u3002\r\n                        <br><i class = \"ChIP-note\">\u8fd9\u91cc\u6c2f\u5316\u9499\u52a0\u5b8c\u4f1a\u5f62\u6210\u4e00\u5c0f\u5c42\u7ec6\u5bc6\u7684\u767d\u8272\u6c89\u6dc0\uff0c\u4f46\u5f88\u5bb9\u6613\u6df7\u5300\uff0c\u800c\u4e14\u6df7\u5300\u540e\u4e0d\u4f1a\u518d\u6c89\u4e0b\u6765\u3002<\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        \u52a0\u5165 <strong style=\"color: green;\">4 \u00b5L<\/strong> \u6d53\u5ea6\u4e3a <strong style=\"color: blue;\">20 mg\/mL<\/strong> \u7684Proteinase K\uff0c\u4f7f\u5176\u7ec8\u6d53\u5ea6\u4e3a <strong style=\"color: blue;\">0.2 mg\/mL<\/strong>\uff0c\u5e76Vortex\u6df7\u5408\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728 <strong style=\"color:rgb(255, 105, 59);\">55\u2103<\/strong> \u7684\u6c34\u6d74\u6216\u6052\u6e29\u6df7\u5408\u5668\uff08Eppendorf Thermomixer C\uff09\u4e2d\u5b75\u80b2 <strong style=\"color: #CC1276;\">0.5-2\u5c0f\u65f6<\/strong>\uff0c\u4ee5\u6d88\u5316\u86cb\u767d\u8d28\u3002\r\n                        <br><i class = \"ChIP-note\">\u5f20\u6d9b\u8bf41h\u5c31\u53ef\u4ee5\uff0c\u4ed6\u6bcf\u6b21\u90fd\u7cbe\u51c6\u5b75\u80b21h\u3002<\/i>\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 14 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 14: \u915a-\u6c2f\u4eff\u63d0\u53d6DNA<\/h4>\r\n                <p style = \"padding-inline-start: 40px;\">\u4f7f\u7528heavy phase-lock tubes\u5728<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u8fdb\u884c\u915a-\u6c2f\u4eff\u63d0\u53d6DNA [\u8bf7\u53c2\u7167heavy phase-lock tubes\u7684protocol]\u3002<\/p>\r\n                <ol>\r\n                    <li>\u6309\u7167\u8bf4\u660e\u79bb\u5fc3 phase-lock tubes\uff08<strong style=\"color: #CC1276;\">13200 rpm<\/strong>\u5ba4\u6e29\u79bb\u5fc3<strong style=\"color: #CC1276;\">30s<\/strong>\uff09\uff0c\u5411\u6bcf\u4e2a\u7ba1\u4e2d\u52a0\u5165 <strong style=\"color: green;\">400 \u00b5L<\/strong> \u6837\u54c1\u3002<\/li>\r\n                    <li>\u52a0\u5165 <strong style=\"color: green;\">1\u500d\u4f53\u79ef\uff08400 \u00b5L\uff09<\/strong> 25:24:1\u7684\u915a-\u6c2f\u4eff-\u5f02\u620a\u9187\uff0c\u731b\u70c8\u6447\u52a8\u81f3\u5c11<strong style=\"color: #CC1276;\">50\u6b21<\/strong>\u6df7\u5300\u3002\r\n                    <i class = \"ChIP-note\"><br>\u6ce8\u610f\u662fDNA\u7684\u915a\u6c2f\u4eff\u5f02\u620a\u9187\uff08pH\uff1e7.8\uff09\uff0c\u8fd8\u6709\u4e00\u79cdpH\uff1c5\u7684\u662f\u7528\u4e8eRNA\u7684\u3002<\/i><\/li>\r\n                    <li>\u4ee5 <strong style=\"color: #CC1276;\">13200 rpm<\/strong> \u7684\u901f\u5ea6\u5728<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u79bb\u5fc3 <strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong> \u4ee5\u5206\u79bb\u5c42\u6b21\u3002<\/li>\r\n                    <li>\u5c06 <strong style=\"color: green;\">400 \u00b5L<\/strong> \u6c2f\u4eff\u52a0\u5165\u6c34\u76f8\uff0c\u731b\u70c8\u6447\u52a8<strong style=\"color: #CC1276;\">50\u6b21<\/strong>\u3002<\/li>\r\n                    <li>\u518d\u6b21\u4ee5 <strong style=\"color: #CC1276;\">13200 rpm<\/strong> \u7684\u901f\u5ea6\u5728<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u79bb\u5fc3 <strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\u3002<\/li>\r\n                    <li>\r\n                        \u5c06\u4e0a\u5c42\u6c34\u76f8\u8f6c\u79fb\u81f3\u65b0\u7684\u4f4e\u5438\u9644\u79bb\u5fc3\u7ba1\u4e2d\u3002\r\n                        <i class = \"ChIP-note\"><br>\u8fd9\u4e24\u6b65\u5267\u70c8\u9707\u8361\u5b8c\u90fd\u4f1a\u770b\u5230\u5927\u91cf\u767d\u8272\u6ce1\u6cab\uff0c\u79bb\u5fc3\u540e\u53d8\u6f84\u6e05\u3002\u52a0\u915a\u6c2f\u4eff\u5f02\u620a\u9187\u548c\u6c2f\u4eff\u7684\u65f6\u5019\uff0c\u90fd\u9700\u8981\u5148\u5438\u4e00\u67aa\u518d\u5410\u56de\u53bb\u6da6\u4e00\u4e0b\u67aa\u5934\uff0c\u4e0d\u7136\u4f1a\u81ea\u52a8\u5f80\u4e0b\u6ef4\uff0c\u53ef\u80fd\u5bfc\u81f4\u52a0\u6837\u4f53\u79ef\u4e0d\u51c6\u3002\u6da6\u8fc7\u4e4b\u540e\u5c31\u4e0d\u5f80\u4e0b\u6ef4\u4e86\u3002<\/i>\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 15 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 15: \u4e59\u9187\u6c89\u6dc0DNA<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u5728\u51b0\u4e0a\u8fdb\u884cDNA\u7684\u4e59\u9187\u6c89\u6dc0 [\u53c2\u9605\u4f34\u968f\u534f\u8bae]\u3002<strong style=\"color:red;\">\u6309\u4ee5\u4e0b\u987a\u5e8f<\/strong>\u6dfb\u52a0\u8bd5\u5242\uff1a\r\n                        <ol class = \"sub-list\">\r\n                            <li>\u52a0\u5165 <strong style=\"color: green;\">1.5 \u00b5L<\/strong> \u6d53\u5ea6\u4e3a <strong style=\"color: blue;\">20 mg\/mL<\/strong> \u7684Glycogen\u3002<\/li>\r\n                            <li>\u52a0\u5165 <strong style=\"color: green;\">1\/10\u4f53\u79ef\uff08\u7ea640 \u00b5L\uff09<\/strong> 3M NaOAc\uff0cVortex\u6df7\u5408\u5e76\u5feb\u901f\u79bb\u5fc3\u3002<\/li>\r\n                            <li>\u52a0\u5165 <strong style=\"color: green;\">2.7\u500d\u4f53\u79ef\uff08\u7ea61080 \u00b5L\uff09<\/strong> \u51b0\u51b7\u7684100%\u4e59\u9187\uff0c\u98a0\u5012<strong style=\"color: #CC1276;\">15\u6b21<\/strong>\u6df7\u5300\u3002<\/li>\r\n                        <\/ol>\r\n                        <i class = \"ChIP-note\"><br>\u603b\u4f53\u79ef\u521a\u597d1521.5\u03bcL\uff0c\u5b9e\u9645\u4e0a\u6837\u54c1\u5438\u4e0d\u51fa\u6765400\u03bcL\u3002\u6709\u4eba\u559c\u6b22\u52a03\u500d\u4f53\u79ef\u4e59\u9187\uff0c\u8fd9\u91cc\u4e00\u5b9a\u5f97\u662f2.7\u500d\uff0c\u9664\u975e\u4f60\u75282mL\u7684\u79bb\u5fc3\u7ba1\u3002\u7cd6\u539f\u548cNaAc\u53ef\u4ee5\u9884\u6df7\uff0c\u4f46\u662f\u4e59\u9187\u4e00\u5b9a\u8981\u6700\u540e\u52a0\u3002<\/i>\r\n                    <\/li>\r\n                    <li>\r\n                        <del style=\"text-decoration-thickness: 2px; background-color: rgb(192, 190, 190);\">\u5728<strong style=\"color:blue;\">-80\u2103<\/strong> \u5b75\u80b2 <strong style=\"color: #CC1276;\">1\u5c0f\u65f6<\/strong> \u6216<\/del>\u5728<strong style=\"color:blue;\">-20\u2103<\/strong> \u8fc7\u591c\u5b75\u80b2\uff0c\u4ee5\u6c89\u6dc0DNA\u3002\r\n                        <i class = \"ChIP-note\"><br>\u5c3d\u91cf\u522b\u7528-80\u2103 1h\uff0c\u6700\u597d\u662f-20\u2103\u8fc7\u591c\u3002\u4e5f\u522b-80\u2103\u8fc7\u591c\uff0c\u56e0\u4e3a-80\u2103\u4f1a\u7ed3\u51b0\u53ef\u80fd\u5f71\u54cd\u9187\u6c89\u6548\u679c\u3002<\/i>\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n        <\/div>\r\n        <div id=\"day4\" class = \"ChIP-day\">\r\n            <h3 >Day4<\/h3>\r\n            <!-- Step 16 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 16: \u79bb\u5fc3\u548c\u6d17\u6da4DNA\u6c89\u6dc0<\/h4>\r\n                <ol>\r\n                    <li>\r\n                        \u4ee5 <strong style=\"color: #CC1276;\">13200 rpm<\/strong> \u7684\u901f\u5ea6\u5728 <strong style=\"color:blue;\">4\u00b0C<\/strong> \u4e0b\u79bb\u5fc3 <strong style=\"color: #CC1276;\">45\u5206\u949f<\/strong>\uff0c\u4ee5\u6c89\u6dc0DNA\uff08\u53ef\u89c1\u5c0f\u7684\u767d\u8272\u6c89\u6dc0\uff09\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u4f7f\u7528\u5c0f\u67aa\u5934\u5c0f\u5fc3\u53bb\u9664\u4e0a\u6e05\u6db2\uff0c\u53ea\u7559\u6c89\u6dc0\u7269\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u52a0\u5165 <strong style=\"color: green;\">0.5 mL<\/strong> \u51b0\u51b7\u7684<strong style=\"color:blue;\">75%\u4e59\u9187<\/strong>\uff0c\u7ffb\u8f6c\u6216\u5f39\u51fb\u4ee5\u91cd\u60ac\uff08\u786e\u4fdd\u6c89\u6dc0\u4e0d\u7c98\u9644\u5728\u6eb6\u6db2\u4e0a\u65b9\u7684\u4efb\u4f55\u533a\u57df\uff0c\u5426\u5219\u96be\u4ee5\u518d\u6b21\u6c89\u6dc0\u81f3\u7ba1\u5e95\uff09\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u4ee5 <strong style=\"color: #CC1276;\">13200 rpm<\/strong> \u7684\u901f\u5ea6\u5728 <strong style=\"color:blue;\">4\u00b0C<\/strong> \u4e0b\u79bb\u5fc3 <strong style=\"color: #CC1276;\">15\u5206\u949f<\/strong>\uff0c\u518d\u6b21\u6c89\u6dc0DNA\uff08\u5c0f\u7684\u767d\u8272\u6c89\u6dc0\uff09\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5438\u5f03\u4e59\u9187\uff0c\u5e76\u4f7f\u7528\u5c0f\u67aa\u5934\u5c0f\u5fc3\u53bb\u9664\u5269\u4f59\u7684\u6db2\u4f53\uff0c\u53ea\u7559\u6c89\u6dc0\u7269\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5728<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong>\u4e0b\u7a7a\u6c14\u5e72\u71e5\u6c89\u6dc0\u7ea6<strong style=\"color: #CC1276;\">10\u5206\u949f<\/strong>\uff0c\u76f4\u5230\u6c89\u6dc0\u53d8\u4e3a\u900f\u660e\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n            <\/div>\r\n\r\n            <!-- Step 17 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 17: \u91cd\u60acDNA\u6c89\u6dc0<\/h4>\r\n                <p style = \"padding-inline-start: 40px;\">\u5c06\u6c89\u6dc0\u91cd\u60ac\u5728\u6240\u9700\u4f53\u79ef\u7684\u65e0\u83cc\u84b8\u998f\u6c34\uff08<strong>dH<sub>2<\/sub>O<\/strong>\uff09\u4e2d\uff0c\u653e\u5165\u6052\u6e29\u6df7\u5408\u5668\uff08Eppendorf Thermomixer C\uff09\u4e2d\uff0c\u5728 <strong style=\"color:orange;\">37\u00b0C<\/strong> \u4e0b\u4ee5 <strong style=\"color: #CC1276;\">1100 rpm<\/strong> \u7684\u901f\u5ea6\u6eb6\u89e3 <strong style=\"color: #CC1276;\">1\u5206\u949f<\/strong>\uff1a\r\n                <br>input = <strong style=\"color: green;\">25 \u00b5L<\/strong>\uff0cChIP\u6837\u54c1 = <strong style=\"color: green;\">25 \u00b5L<\/strong><\/p>\r\n            <\/div>\r\n\r\n            <!-- Step 18 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 18: \u6d4b\u5b9aDNA\u6d53\u5ea6<\/h4>\r\n                <p style = \"padding-inline-start: 40px;\">\u901a\u8fc7Qubit\u6d4b\u5b9aDNA\u7684\u6d53\u5ea6\u3002<\/p>\r\n                <i class = \"ChIP-note\"><br>\u6309\u7167\u7ecf\u9a8c\uff0cInput\u5927\u6982\u4e2a\u4f4d\u6570ng\uff0cIP\u7684Elute\u5927\u69820.1~0.2ng<\/i>\r\n            <\/div>\r\n\r\n            <!-- Step 19 -->\r\n            <div class = \"ChIP-step\">\r\n                <h4>\u6b65\u9aa4 19: \u6784\u5efa\u6587\u5e93\u6216\u4fdd\u5b58DNA<\/h4>\r\n                <p style = \"padding-inline-start: 40px;\">\u6784\u5efa\u6d4b\u5e8f\u6587\u5e93\uff0c\u6216\u8005\u5c06DNA\u5b58\u50a8\u5728 <strong style=\"color:blue;\">-20\u00b0C<\/strong> \u76f4\u5230\u9700\u8981\u8fdb\u884c\u540e\u7eed\u6b65\u9aa4\u3002<\/p>\r\n                <i class = \"ChIP-note\"><br>\u8fd9\u91cc\u7528ABclonal\u7684Rapid Plus DNA Lib Prep Kit (RK20208)<\/i>\r\n\r\n                <h5>1. \u672b\u7aef\u4fee\u590d<\/h5>\r\n                <ol>\r\n                    <li>\r\n                        \u51c6\u5907\u516b\u8054\u6392\u7ba1\uff0c\u6bcf\u7ba1\u5185\u9884\u6df7 <strong>End Prep Buffer<\/strong> <strong style=\"color:green;\">3.5\u03bcL<\/strong> + <strong>End Prep Enzymes<\/strong> <strong style=\"color:green;\">1.5\u03bcL<\/strong> + <strong>Damaged DNA Repair Enzymes<\/strong> <strong style=\"color:green;\">1.5\u03bcL<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5bf9\u4e8e<strong>Input<\/strong>\u6837\u54c1\uff0c\u53d6<strong style=\"color: purple;\">30ng<\/strong> \u52a0\u5165\u4e0a\u8ff0\u4f53\u7cfb\uff0c\u518d\u7528\u6c34\u8865\u8db3\u81f3\u603b\u4f53\u79ef <strong style=\"color: green;\">30\u03bcL<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5bf9\u4e8e\u5176\u4ed6<strong>Elute<\/strong>\u6837\u54c1\uff0c\u53d6 <strong style=\"color: green;\">23.5\u03bcL<\/strong> \u52a0\u5165\u4e0a\u8ff0\u4f53\u7cfb\uff0c\u603b\u4f53\u79ef\u540c\u6837\u4e3a <strong style=\"color: green;\">30\u03bcL<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u8fdb\u884c\u672b\u7aef\u4fee\u590d\u7a0b\u5e8f\uff1a<strong style=\"color: #76a802;\">30\u2103<\/strong>, <span style=\"color: #CC1276;\">30min<\/span> \u2192 <strong style=\"color: orange;\">65\u2103<\/strong>, <span style=\"color: #CC1276;\">30min<\/span> \u2192 <strong style=\"color: blue;\">4\u2103<\/strong> , \u221e\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n\r\n                <h5>2. \u63a5\u5934\u8fde\u63a5<\/h5>\r\n                 <ol>\r\n                    <li>\r\n                        \u6839\u636e\u6295\u5165DNA\u7684\u8d28\u91cf\u6765\u51b3\u5b9a\u63a5\u5934\u7684\u7a00\u91ca\u500d\u6570\uff1a\r\n                        <ul>\r\n                            <li>\r\n                                \u5bf9\u4e8e <strong style=\"color: purple;\">30ng<\/strong> <strong>Input<\/strong>\uff0c\u7528 <strong style=\"color: blue;\">Low-EDTA TE<\/strong> \u7a00\u91ca <strong style=\"color: #CC1276;\">2\u500d<\/strong>\u3002<br>\r\n                            <\/li>\r\n                            <li>\r\n                                \u5bf9\u4e8e IP \u7684 <strong>Elute<\/strong>\uff0c\u6d53\u5ea6\u5728 <strong style=\"color: blue;\">42.6 ng\/\u00b5L ~ 2.1 ng\/\u00b5L<\/strong> \u8303\u56f4\u5185\u7684\u65e0\u9700\u7a00\u91ca\uff1b<strong style=\"color: blue;\">2.1 ng\/\u00b5L ~ 1.1 ng\/\u00b5L<\/strong> \u7a00\u91ca <strong style=\"color: #CC1276;\">2\u500d<\/strong>\uff1b<strong style=\"color: blue;\">1.0 ng\/\u00b5L ~ 0.4 ng\/\u00b5L<\/strong> \u7a00\u91ca <strong style=\"color: #CC1276;\">5\u500d<\/strong>\uff1b<strong style=\"color: blue;\">0.4 ng\/\u00b5L ~ 0.2 ng\/\u00b5L<\/strong> \u7a00\u91ca <strong style=\"color: #CC1276;\">10\u500d<\/strong>\uff1b<strong style=\"color: blue;\">< 0.2 ng\/\u00b5L<\/strong> \u7a00\u91ca <strong style=\"color: #CC1276;\">20\u500d<\/strong>\u3002\r\n                            <\/li>\r\n                        <\/ul>\r\n                    <\/li>\r\n                    <li>\r\n                        \u51c6\u5907\u63a5\u5934\u8fde\u63a5\u53cd\u5e94\u4f53\u7cfb\uff08\u5355\u4e2a\u6837\u672c\uff09\uff1a<strong>End Prep Reaction Mix<\/strong> <strong style=\"color: green;\">30 \u00b5L<\/strong>\uff0c<strong>Ligation Buffer<\/strong> <strong style=\"color: green;\">15 \u00b5L<\/strong>\uff0c<strong>ddH\u2082O<\/strong> <strong style=\"color: green;\">2.5 \u00b5L<\/strong>\uff0c<strong>Ligation Enzymes<\/strong> <strong style=\"color: green;\">5 \u00b5L<\/strong>\uff0c<strong>Working Adaptor<\/strong> <strong style=\"color: green;\">2.5 \u00b5L<\/strong>\uff0c\u603b\u4f53\u79ef <strong style=\"color: green;\">55 \u00b5L<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u53cd\u5e94\u7a0b\u5e8f\u4e3a <strong style=\"color: blue;\">22\u2103<\/strong>\uff0c<strong style = \"color:#CC1276;\">15 \u5206\u949f<\/strong>\uff0cPCR \u4eea\u4e0d\u8bbe\u70ed\u76d6\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n\r\n                <h5>3. \u78c1\u73e0\u7eaf\u5316<\/h5>\r\n                <ol>\r\n                    <li>\r\n                        \u6bcf\u4e2a\u6837\u672c\u4e2d\u52a0\u5165 <strong style=\"color: green;\">44\u03bcL (0.8X)<\/strong> Agencourt\u2122 Ampure XP Beads \u5e76\u6df7\u5300\u540e\uff0c\u5728 <strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong> \u4e0b\u5b75\u80b2 <strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5c06PCR\u7ba1\u653e\u7f6e\u4e8e\u78c1\u529b\u67b6\u4e0a\u9759\u7f6e <strong style=\"color: #CC1276;\">2\u5206\u949f<\/strong>\uff0c\u5f85\u6eb6\u6db2\u6f84\u6e05\u540e\u5c0f\u5fc3\u79fb\u9664\u4e0a\u6e05\u6db2\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u4f7f\u7528 <strong style=\"color: green;\">100\u03bcL<\/strong><strong> 80%\u4e59\u9187<\/strong>\u6f02\u6d17\u78c1\u73e0\uff0c\u5b75\u80b2 <strong style=\"color: #CC1276;\">30\u79d2<\/strong> \u540e\u518d\u79fb\u9664\u4e0a\u6e05\uff0c\u5e76\u91cd\u590d\u6b64\u6f02\u6d17\u6b65\u9aa4\u4e00\u6b21\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u4fdd\u6301PCR\u7ba1\u5728\u78c1\u529b\u67b6\u4e0a\uff0c\u7528 <strong style=\"color: green;\">10\u03bcL<\/strong> \u79fb\u6db2\u5668\u5438\u8d70\u7ba1\u5e95\u6b8b\u7559\u7684\u4e59\u9187\uff0c\u5e76\u6253\u5f00\u7ba1\u76d6\u4f7f\u78c1\u73e0\u5e72\u71e5\u76f4\u81f3\u65e0\u4e59\u9187\u6b8b\u7559\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u6700\u540e\uff0c\u5728 <strong style=\"color: green;\">10.5\u03bcL<\/strong>  <strong>Low-EDTA TE Buffer<\/strong>\u4e2d\u91cd\u60ac\u78c1\u73e0\uff0c<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong> \u9759\u7f6e <strong style=\"color: #CC1276;\">1\u5206\u949f<\/strong> \u540e\uff0c\u518d\u6b21\u7f6e\u4e8e\u78c1\u529b\u67b6\u4e0a\u9759\u7f6e <strong style=\"color: #CC1276;\">2\u5206\u949f<\/strong>\uff0c\u7136\u540e\u8f6c\u79fb <strong style=\"color: green;\">10\u03bcL<\/strong> \u4e0a\u6e05\u6db2\u81f3\u65b0\u7684PCR\u7ba1\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n\r\n                <h5>4. PCR\u6269\u589e<\/h5>\r\n                <ol>\r\n                    <li>\r\n                        \u51c6\u5907PCR\u53cd\u5e94\u6df7\u5408\u7269\uff0c\u5305\u542b <strong style=\"color: green;\">5 \u03bcL<\/strong> <strong>\u63a5\u5934\u8fde\u63a5\u7684DNA<\/strong>\u3001<strong style=\"color: green;\">6.25 \u03bcL<\/strong> <strong>2X PCR Master Mix<\/strong> \u548c <strong style=\"color: green;\">1.25 \u03bcL<\/strong> <strong>Dual Index Primers<\/strong>\uff0c\u603b\u4f53\u79ef <strong style=\"color: green;\">12.5 \u03bcL<\/strong>\u3002                        \r\n                    <\/li>\r\n                    <li>\r\n                        \u8fdb\u884c\u4ee5\u4e0b\u70ed\u5faa\u73af\uff1a<strong style=\"color: red;\">98\u2103<\/strong>\u9884\u53d8\u6027<strong style=\"color: #CC1276;\">1\u5206\u949f<\/strong>\uff1b\u968f\u540e\u662f <strong style=\"color: red;\">98\u2103<\/strong> <strong style=\"color: #CC1276;\">10\u79d2<\/strong>\u3001<strong style=\"color: orange;\">60\u2103<\/strong> <strong style=\"color: #CC1276;\">30\u79d2<\/strong>\u3001<strong style=\"color: #fd8003;\">72\u2103<\/strong> <strong style=\"color: #CC1276;\">30\u79d2<\/strong>\uff0c\u51712\u81f317\u4e2a\u5faa\u73af\uff08Input <strong>5\u4e2a\u5faa\u73af<\/strong>\uff0cElute <strong>12\u4e2a\u5faa\u73af<\/strong>\uff09\uff1b\u6700\u540e <strong style=\"color: #fd8003;\">72\u2103<\/strong>\u5ef6\u4f38<strong style=\"color: #CC1276;\">1\u5206\u949f<\/strong>\uff0c\u4fdd\u6301\u5728 <strong style=\"color: blue;\">12\u2103<\/strong>\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n                <ol>\r\n                    <li>\r\n                        \u51c6\u59071%\u743c\u8102\u7cd6\u51dd\u80f6\uff0c\u53d6<strong style = \"color: green;\">1\u03bcL<\/strong>PCR\u4ea7\u7269\u7535\u6cf3\u68c0\u6d4bPCR\u7ed3\u679c\uff0c\u5bf9\u4ea7\u91cf\u4e0d\u8db3\u7684\u6837\u54c1\u589e\u52a0PCR\u5faa\u73af\u6b21\u6570\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u52a0\u5b8c\u5faa\u73af\u6570\u540e\u518d\u53d61\u03bcL\u8dd1\u80f6\u786e\u8ba4\u4ea7\u91cf\u3002\r\n                    <\/li>\r\n                <\/ol>\r\n                <h5>5. \u7b2c\u4e8c\u6b21\u78c1\u73e0\u7eaf\u5316<\/h5>\r\n                <ol>\r\n                    <li>\r\n                        \u6bcf\u4e2a\u6837\u672c\u4e2d\u52a0\u5165 <strong style=\"color: green;\">10.5\u03bcL \/ 11.5\u03bcL (1X)<\/strong> Agencourt\u2122 Ampure XP Beads \u5e76\u6df7\u5300\u540e\uff0c\u5728 <strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong> \u4e0b\u5b75\u80b2 <strong style=\"color: #CC1276;\">5\u5206\u949f<\/strong>\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u5c06PCR\u7ba1\u653e\u7f6e\u4e8e\u78c1\u529b\u67b6\u4e0a\u9759\u7f6e <strong style=\"color: #CC1276;\">2\u5206\u949f<\/strong>\uff0c\u5f85\u6eb6\u6db2\u6f84\u6e05\u540e\u5c0f\u5fc3\u79fb\u9664\u4e0a\u6e05\u6db2\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u4f7f\u7528 <strong style=\"color: green;\">100\u03bcL<\/strong> <strong>80%\u4e59\u9187<\/strong> \u6f02\u6d17\u78c1\u73e0\uff0c\u5b75\u80b2 <strong style=\"color: #CC1276;\">30\u79d2<\/strong> \u540e\u518d\u79fb\u9664\u4e0a\u6e05\uff0c\u5e76\u91cd\u590d\u6b64\u6f02\u6d17\u6b65\u9aa4\u4e00\u6b21\u3002\r\n                    <\/li>\r\n                    <li>\r\n                        \u4fdd\u6301PCR\u7ba1\u5728\u78c1\u529b\u67b6\u4e0a\uff0c\u752810\u03bcL\u79fb\u6db2\u5668\u5438\u8d70\u7ba1\u5e95\u6b8b\u7559\u7684\u4e59\u9187\uff0c\u5e76\u6253\u5f00\u7ba1\u76d6\u4f7f\u78c1\u73e0\u5e72\u71e5\u76f4\u81f3\u65e0\u4e59\u9187\u6b8b\u7559\u3002\r\n                        <\/li>\r\n                        <li>\r\n                            \u6700\u540e\uff0c\u5728 <strong style=\"color: green;\">15\u03bcL <\/strong><strong>Low-EDTA TE Buffer<\/strong> \u4e2d\u91cd\u60ac\u78c1\u73e0\uff0c<strong style=\"color: rgb(0, 150, 150);\">\u5ba4\u6e29<\/strong> \u9759\u7f6e <strong style=\"color: #CC1276;\">1\u5206\u949f<\/strong> \u540e\uff0c\u518d\u6b21\u7f6e\u4e8e\u78c1\u529b\u67b6\u4e0a\u9759\u7f6e <strong style=\"color: #CC1276;\">2\u5206\u949f<\/strong>\uff0c\u7136\u540e\u8f6c\u79fb <strong style=\"color: green;\">15\u03bcL<\/strong> \u4e0a\u6e05\u6db2\u81f3\u65b0\u7684PCR\u7ba1\u3002\r\n                        <\/li>\r\n                    <\/ol>\r\n\r\n                    <h5>7. Qubit\u5b9a\u91cf<\/h5>\r\n                    <p>\u5bf9\u7ecf\u8fc7\u7b2c\u4e8c\u6b21\u78c1\u73e0\u7eaf\u5316\u7684\u6837\u54c1\u8fdb\u884cQubit\u5b9a\u91cf\uff0c\u786e\u4fdd\u5176\u6d53\u5ea6\u7b26\u5408\u540e\u7eed\u5b9e\u9a8c\u8981\u6c42\u3002<\/p>\r\n                <\/div>\r\n            <\/div>\r\n        <\/div>\r\n    <div id=\"reagent\" class = \"ChIP-day\">\r\n            <h3 >\u9700\u8981\u914d\u7f6e\u7684\u8bd5\u5242<\/h3>\r\n            <h4 style=\"margin-left: 10px;\">2.5M Glycine <span style=\"color: rgb(0, 150, 150);\">[make fresh, store at RT]<\/span> Used in Day1<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Amount<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td>Glycine<\/td>\r\n                        <td><strong style=\"color: green;\">0.94 g<\/strong><\/td>\r\n                        <td>2.5 M Glycine<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>H<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">5 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n        \r\n            <h4 style=\"margin-left: 10px;\">Lysis Buffer 1 \u2013 LB1 (50 mL) <span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span> Used in Day1<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1M<\/span> Hepes-KOH pH 7.5<\/td>\r\n                        <td><strong style=\"color: green;\">2.5 mL<\/strong><\/td>\r\n                        <td>50 mM Hepes<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">5M<\/span> NaCl<\/td>\r\n                        <td><strong style=\"color: green;\">1.4 mL<\/strong><\/td>\r\n                        <td>140 mM NaCl<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">100 \u00b5L<\/strong><\/td>\r\n                        <td>1 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">50% <\/span>Glycerol<\/td>\r\n                        <td><strong style=\"color: green;\">10 mL<\/strong><\/td>\r\n                        <td>10% Glycerol<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">10% <\/span>NP-40<\/td>\r\n                        <td><strong style=\"color: green;\">2.5 mL<\/strong><\/td>\r\n                        <td>0.5% NP-40<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">10% <\/span>Triton X-100 (PBS)<\/td>\r\n                        <td><strong style=\"color: green;\">1.25 mL<\/strong><\/td>\r\n                        <td>0.25% TX-100<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">32.25 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n        \r\n            <h4 style=\"margin-left: 10px;\">Lysis Buffer 2 \u2013 LB2 (50 mL) <span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span> Used in Day1<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1 M<\/span> Tris-HCl pH 8.0<\/td>\r\n                        <td><strong style=\"color: green;\">0.5 mL<\/strong><\/td>\r\n                        <td>10 mM Tris<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">5 M<\/span> NaCl<\/td>\r\n                        <td><strong style=\"color: green;\">2.0 mL<\/strong><\/td>\r\n                        <td>200 mM NaCl<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">100 \u00b5L<\/strong><\/td>\r\n                        <td>1 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EGTA<\/td>\r\n                        <td><strong style=\"color: green;\">50 \u00b5L<\/strong><\/td>\r\n                        <td>0.5 mM EGTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">47.35 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n        \r\n            <h4 style=\"margin-left: 10px;\">Shearing Buffer 1 (10 mL) <span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span> Used in Day1<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1 M<\/span> Tris-HCl pH 7.6<\/td>\r\n                        <td><strong style=\"color: green;\">0.1 mL<\/strong><\/td>\r\n                        <td>10 mM Tris<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">20 \u00b5L<\/strong><\/td>\r\n                        <td>1 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">30%<\/span> SDS<\/td>\r\n                        <td><strong style=\"color: green;\">0.1 mL<\/strong><\/td>\r\n                        <td>0.3% SDS<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">9.78 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n        \r\n            <h4 style=\"margin-left: 10px;\">Shearing Buffer 2 (10 mL) <span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span> Used in Day1<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1 M<\/span> Tris-HCl pH 7.6<\/td>\r\n                        <td><strong style=\"color: green;\">0.1 mL<\/strong><\/td>\r\n                        <td>10 mM Tris<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">20 \u00b5L<\/strong><\/td>\r\n                        <td>1 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">9.88 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n            <h4 style=\"margin-left: 10px;\">Block Solution (50 mL) <span style=\"color: blue;\">[store at 4\u2103] <\/span> Used in Day2\uff08\u73b0\u914d\u73b0\u7528\uff09<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td>BSA powder<\/td>\r\n                        <td><strong style=\"color: green;\">125mg<\/strong><\/td>\r\n                        <td>0.5% BSA (w\/v)<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td> PBS (1x)<\/td>\r\n                        <td><strong style=\"color: green;\">25mL<\/strong><\/td>\r\n                        <td>1x PBS<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n            <h4 style=\"margin-left: 10px;\">RIPA Wash Buffer (250 mL) <span style=\"color: blue;\">[store at 4\u00b0C]<\/span> Used in Day2<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1 M<\/span> Hepes-KOH pH 7.5<\/td>\r\n                        <td><strong style=\"color: green;\">12.5 mL<\/strong><\/td>\r\n                        <td>50 mM Hepes<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">8 M<\/span> LiCl<\/td>\r\n                        <td><strong style=\"color: green;\">15.6 mL<\/strong><\/td>\r\n                        <td>500 mM LiCl<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">0.5 mL<\/strong><\/td>\r\n                        <td>1 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">10%<\/span> NP-40<\/td>\r\n                        <td><strong style=\"color: green;\">25.0 mL<\/strong><\/td>\r\n                        <td>1% NP-40<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">10%<\/span> Na-Deoxycholate<\/td>\r\n                        <td><strong style=\"color: green;\">17.5 mL<\/strong><\/td>\r\n                        <td>0.7% Na-Deoxycholate<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">178.9 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n\r\n            <h4 style=\"margin-left: 10px;\">Elution Buffer (50 mL) <span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span><\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1 M<\/span> Tris-HCl pH 8.0<\/td>\r\n                        <td><strong style=\"color: green;\">2.5 mL<\/strong><\/td>\r\n                        <td>50 mM Tris<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">1.0 mL<\/strong><\/td>\r\n                        <td>10 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">10%<\/span> SDS<\/td>\r\n                        <td><strong style=\"color: green;\">5.0 mL<\/strong><\/td>\r\n                        <td>1% SDS<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">41.5 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n\r\n            <h4 style=\"margin-left: 10px;\">1X TE Buffer, pH 8.0 \uff0850mL\uff09<span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span><\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">1 M<\/span> Tris-HCl pH 8.0<\/td>\r\n                        <td><strong style=\"color: green;\">0.5 mL<\/strong><\/td>\r\n                        <td>10 mM Tris-HCl<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td><span style=\"color: blue;\">0.5 M<\/span> EDTA<\/td>\r\n                        <td><strong style=\"color: green;\">0.1 mL<\/strong><\/td>\r\n                        <td>1 mM EDTA<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">49.4 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n            <h4 style=\"margin-left: 10px;\">300 mM CaCl\u2082 (10 mL) <span style=\"color: rgb(0, 150, 150);\">[store at RT]<\/span> Used in Day3<\/h4>\r\n            <table border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n                <thead>\r\n                    <tr>\r\n                        <th style=\"background-color: #f0f8ff;\">Component<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Volume<\/th>\r\n                        <th style=\"background-color: #f0f8ff;\">Final Concentration<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr>\r\n                        <td>CaCl\u2082<\/td>\r\n                        <td><strong style=\"color: green;\">0.333 g<\/strong><\/td>\r\n                        <td>300 mM CaCl\u2082<\/td>\r\n                    <\/tr>\r\n                    <tr>\r\n                        <td>dH<sub>2<\/sub>O<\/td>\r\n                        <td><strong style=\"color: green;\">to 10 mL<\/strong><\/td>\r\n                        <td>-<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n\r\n            <h3 >\u90e8\u5206\u50a8\u6db2\u914d\u65b9<\/h3>\r\n\r\n            <table class=\"Stock-table\">\r\n                <thead>\r\n                    <tr class=\"Stock-tr\">\r\n                        <th class=\"Stock-th\">\u914d\u65b9<\/th>\r\n                        <th class=\"Stock-th\">\u914d\u5236\u65b9\u6cd5<\/th>\r\n                        <th class=\"Stock-th\">\u4f53\u79ef<\/th>\r\n                        <th class=\"Stock-th\">\u50a8\u85cf\u6e29\u5ea6<\/th>\r\n                        <th class=\"Stock-th\">\u6ce8\u610f\u4e8b\u9879<\/th>\r\n                    <\/tr>\r\n                <\/thead>\r\n                <tbody>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">5M NaCl<\/td>\r\n                        <td class=\"Stock-td\">14.61g NaCl \u52a0\u6c34\u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u9632\u6b62\u7ed3\u6676\uff0c\u5bc6\u5c01\u4fdd\u5b58\u3002\u6eb6\u89e3\u8f83\u6162\uff0c\u9700\u8981\u7528\u5230\u65f6\u63d0\u524d\u7559\u51fa\u65f6\u95f4\u914d\u7f6e\u3002<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">0.5M EDTA<\/td>\r\n                        <td class=\"Stock-td\">2.79g EDTA \u52a0\u6c34\u81f3 15mL\uff0c\u8c03\u8282pH\u81f38.0<\/td>\r\n                        <td class=\"Stock-td\">15 mL<\/td>\r\n                        <td class=\"Stock-td\">4\u00b0C<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u91d1\u5c5e\u79bb\u5b50\u6c61\u67d3<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">10% NP-40<\/td>\r\n                        <td class=\"Stock-td\">5mL NP-40 \u52a0\u6c34\u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u6ce1\u6cab\uff0c\u5bc6\u5c01\u4fdd\u5b58\u3002\u6eb6\u89e3\u8f83\u6162\uff0c\u9700\u8981\u7528\u5230\u65f6\u63d0\u524d\u7559\u51fa\u65f6\u95f4\u914d\u7f6e\u3002<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">50% \u7518\u6cb9<\/td>\r\n                        <td class=\"Stock-td\">25mL \u7518\u6cb9 \u52a0\u6c34\u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u5bc6\u5c01\u4fdd\u5b58\uff0c\u907f\u514d\u5438\u6e7f<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">10% Triton X-100 (in PBS)<\/td>\r\n                        <td class=\"Stock-td\">5mL Triton X-100 \u52a0 PBS \u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u6ce1\u6cab\uff0c\u8f7b\u67d4\u6df7\u5300<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">0.5M EGTA<\/td>\r\n                        <td class=\"Stock-td\">1.42g EGTA \u52a0\u6c34\u81f3 15mL\uff0c\u8c03\u8282pH\u81f38.0<\/td>\r\n                        <td class=\"Stock-td\">15 mL<\/td>\r\n                        <td class=\"Stock-td\">4\u00b0C<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u91d1\u5c5e\u79bb\u5b50\u6c61\u67d3<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">30% SDS<\/td>\r\n                        <td class=\"Stock-td\">15g SDS \u52a0\u6c34\u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u5438\u6e7f\uff0c\u5bc6\u5c01\u4fdd\u5b58<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">10% SDS<\/td>\r\n                        <td class=\"Stock-td\">5g SDS \u52a0\u6c34\u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u5438\u6e7f\uff0c\u5bc6\u5c01\u4fdd\u5b58<\/td>\r\n                    <\/tr>\r\n                    <tr class=\"Stock-tr\">\r\n                        <td class=\"Stock-td\">10% Na-Deoxycholate<\/td>\r\n                        <td class=\"Stock-td\">5g Na-Deoxycholate \u52a0\u6c34\u81f3 50mL<\/td>\r\n                        <td class=\"Stock-td\">50 mL<\/td>\r\n                        <td class=\"Stock-td\">\u5ba4\u6e29<\/td>\r\n                        <td class=\"Stock-td\">\u907f\u514d\u6ce1\u6cab\uff0c\u5bc6\u5c01\u4fdd\u5b58\u3002\u6eb6\u89e3\u8f83\u6162\uff0c\u9700\u8981\u7528\u5230\u65f6\u63d0\u524d\u7559\u51fa\u65f6\u95f4\u914d\u7f6e\u3002<\/td>\r\n                    <\/tr>\r\n                <\/tbody>\r\n            <\/table>\r\n        <\/div>\r\n    <div id=\"reagentCalc\">\r\n        <h3>\u5b9e\u9a8c\u6240\u9700\u8bd5\u5242\u53ca\u7528\u91cf<\/h3>\r\n        <label for=\"sampleCount\">\u8bf7\u8f93\u5165\u6837\u54c1\u6570\u91cf (n): <\/label>\r\n        <input type=\"number\" id=\"sampleCount\" value=\"1\">\r\n        <table id=\"reagent-table\" border=\"1\" cellspacing=\"0\" cellpadding=\"5\">\r\n            <thead style=\"background-color: #f0f0f0; font-weight: bold; font-size: 14px;\">\r\n                <tr>\r\n                    <th style=\"background-color: #f0f8ff;\">\u8bd5\u5242\u540d\u79f0<\/th>\r\n                    <th style=\"background-color: #f0f8ff;\">\u7528\u9014<\/th>\r\n                    <th style=\"background-color: #f0f8ff;\">\u5355\u4e2a\u6837\u54c1\u7528\u91cf<\/th>\r\n                    <th style=\"background-color: #f0f8ff;\">\u603b\u7528\u91cf (n)<\/th>\r\n                <\/tr>\r\n            <\/thead>\r\n            <tbody id=\"reagentTableBody\" style=\"font-family: 'Arial', sans-serif; font-size: 14px; line-height: 1.5;\">\r\n                <!-- \u52a8\u6001\u751f\u6210\u884c -->\r\n            <\/tbody>\r\n        <\/table>\r\n        \r\n    <\/div>\r\n\r\n    <!-- \u6dfb\u52a0\u6309\u94ae -->\r\n    <div id=\"day-buttons\" style=\"padding-left: 10px;\">\r\n        <button class=\"ChIP-button\" onclick=\"showDay('day1')\">Day 1<\/button>\r\n        <button class=\"ChIP-button\" onclick=\"showDay('day2')\">Day 2<\/button>\r\n        <button class=\"ChIP-button\" onclick=\"showDay('day3')\">Day 3<\/button>\r\n        <button class=\"ChIP-button\" onclick=\"showDay('day4')\">Day 4<\/button>\r\n        <button class=\"ChIP-button\" onclick=\"showDay('reagent')\">reagent<\/button>\r\n        <button class=\"ChIP-button\" onclick=\"showDay('reagentCalc')\">reagentCalc<\/button>\r\n    <\/div>\n\n\n\n<style>\n#Experiment-Process .container {padding-left: 0; padding-right: 0;}\n#Experiment-Process h4 {margin-top: 0;}\n#Experiment-Process .ChIP-step p {padding-inline-start: 0 !important;}\n<\/style>\n\n\n\n<p><\/p>\n","protected":false},"excerpt":{"rendered":"<p>\u8fd9\u7bc7\u7b14\u8bb0\u8be6\u7ec6\u8bb0\u5f55\u4e86ChIP-seq\u5b9e\u9a8c\u7684\u5b8c\u6574\u64cd\u4f5c\u6d41\u7a0b\uff0c\u4ece\u7ec6\u80de\u4ea4\u8054\u3001\u67d3\u8272\u8d28\u7247\u6bb5\u5316\u5230\u514d\u75ab\u6c89\u6dc0\u4e0e\u6587\u5e93\u6784\u5efa\u3002\u5b83\u7cfb\u7edf\u68b3\u7406\u4e86\u6bcf\u4e2a\u6b65\u9aa4\u7684\u5173\u952e\u53c2\u6570\u3001\u8bd5\u5242\u9009\u62e9\u4e0e\u5e38\u89c1\u95ee\u9898\uff0c\u4e3a\u8868\u89c2\u9057\u4f20\u5b66\u7814\u7a76\u63d0\u4f9b\u4e86\u5b9e\u7528\u7684\u6280\u672f\u53c2\u8003\u3002<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[],"class_list":["post-6758","post","type-post","status-publish","format-standard","hentry","category-note"],"_links":{"self":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/6758","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=6758"}],"version-history":[{"count":19,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/6758\/revisions"}],"predecessor-version":[{"id":13917,"href":"https:\/\/fanyiming.life\/index.php?rest_route=\/wp\/v2\/posts\/6758\/revisions\/13917"}],"wp:attachment":[{"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=6758"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=6758"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/fanyiming.life\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=6758"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}