{"id":8977,"date":"2020-12-02T12:06:59","date_gmt":"2020-12-02T04:06:59","guid":{"rendered":"http:\/\/mp.weixin.qq.com\/s?__biz=MzI3NjUzOTM1Mg==&mid=100017724&idx=1&sn=31d13daae1860a4bd6297651517805e5&chksm=6b71748b5c06fd9d91f6547608ae9f02f1d96d3afa23702776617700b7a821a26ab42e61cda5#rd"},"modified":"2026-04-08T09:35:50","modified_gmt":"2026-04-08T01:35:50","slug":"515051","status":"publish","type":"post","link":"https:\/\/www.geneclub.net.cn\/index.php\/2020\/12\/02\/515051\/","title":{"rendered":"\u7532\u57fa\u5316\u68c0\u6d4b\u4e28\u63a2\u7d22\u6587\u5e93\u6784\u5efa\u4e2d\u66f4\u6e29\u548c\u7684DNA\u5904\u7406\u65b9\u5f0f"},"content":{"rendered":"

<\/p>\n

\u3010\u5bfc\u8bed\u3011\u76ee\u524d\u8f83\u5e38\u4f7f\u7528\u7684DNA\u7532\u57fa\u5316\u68c0\u6d4b\u65b9\u6cd5\u5927\u90fd\u9700\u8981BS\uff08\u91cd\u4e9a\u786b\u9178\u6c22\u76d0\uff09\u5904\u7406\uff0c\u8be5\u6280\u672f\u5bb9\u6613\u5bfc\u81f4\u7edd\u5927\u591a\u6570DNA\u5206\u5b50\u53d1\u751f\u65ad\u94fe\u3002\u672c\u6587\u4e3b\u8981\u5bf9\u7532\u57fa\u5316\u6587\u5e93\u6784\u5efa\u8fc7\u7a0b\u4e2dBS\u5904\u7406\u65b9\u6848\u548c\u9176\u5904\u7406\u65b9\u6848\u8fdb\u884c\u5bf9\u6bd4\uff0c\u4f9b\u65b9\u6848\u8bc4\u4f30\u4e0e\u9009\u62e9\u53c2\u8003\u3002<\/div>\n
\u6587\u7ae0 | \u5143\u7801\u57fa\u56e0\u7f16\u8f91 | \u7f57\u6e58\u00a0 \u5173\u952e\u8bcd | DNA\u7532\u57fa\u5316<\/div>\n
\u00a0<\/div>\n
DNA\u7532\u57fa\u5316<\/div>\n
DNA\u7532\u57fa\u5316\u662fDNA\u5316\u5b66\u4fee\u9970\u7684\u4e00\u79cd\u5f62\u5f0f\uff0c\u80fd\u5728\u4e0d\u6539\u53d8DNA\u5e8f\u5217\u7684\u524d\u63d0\u4e0b\uff0c\u6539\u53d8\u9057\u4f20\u8868\u73b0\u3002<\/div>\n
DNA\u80de\u5627\u5576\u7b2c\u4e94\u4f4d\u7684\u7532\u57fa\u5316\u5728\u591a\u79cd\u751f\u7269\u4e2d\u662f\u7a33\u5b9a\u5b58\u5728\u7684\u4e00\u79cd\u8868\u89c2\u4fee\u9970\uff0c\u5b83\u5df2\u77e5\u5728\u80da\u80ce\u53d1\u80b2\u3001\u8f6c\u5ea7\u5b50\u6c89\u9ed8\u3001X\u67d3\u8272\u4f53\u5931\u6d3b\u3001\u7ec6\u80de\u5206\u5316\u7b49\u591a\u79cd\u8fc7\u7a0b\u4e2d\u901a\u8fc7\u8f6c\u5f55\u6d3b\u6027\u8c03\u8282\u8d77\u5230\u91cd\u8981\u4f5c\u7528\u3002<\/div>\n\n
\n
\"RmFicvQBAkgBCnoXwL82fPGtN82mIqoKus8ZGe6nOydXiao4s0cuibBELY50qkXCmhib8oZDTxEJzZMUeOUpZicoKBA\"\/<\/figure><\/div>\n\n<\/div>\n
\u56fe\uff1aDNA\u7532\u57fa\u5316\uff08\u6765\u6e90\/Google\uff09<\/div>\n
\u81ea1992\u5e74[1]\u9996\u6b21\u4f7f\u7528\u4ee5\u6765\uff0c\u91cd\u4e9a\u786b\u9178\u6c22\u76d0(BS)\u6d4b\u5e8fDNA\u5df2\u6210\u4e3a\u5206\u6790DNA\u7532\u57fa\u5316\u7684\u91d1\u6807\u51c6\u3002\u5bf9DNA\u7684BS\u5904\u7406\u5bfc\u81f4\u672a\u4fee\u9970\u7684\u80de\u5627\u5576\u8f6c\u5316\u4e3a\u5c3f\u5627\u5576\uff0c\u540c\u65f6\u4fdd\u63015mC\u4e0d\u53d8\uff0c\u7ecf\u8fc7PCR\u548c\u6d4b\u5e8f\uff0c\u53ef\u4ee5\u5728\u5355\u78b1\u57fa\u5206\u8fa8\u7387[2,3]\u4e0a\u5b9a\u4f4d\u3002<\/div>\n
\u6700\u8fd1\uff0cBS\u5904\u7406\u548c\u4e8c\u4ee3\u6d4b\u5e8f\u76f8\u7ed3\u5408\u53ef\u4ee5\u5f97\u52305mC\u7684\u5168\u57fa\u56e0\u7ec4\u5206\u5e03\u60c5\u51b5\uff0c\u6bd4\u5982Reduced Representation BS\uff08RRBS\uff09\u6216\u8005Whole Genome BS(WGBS)\u65b9\u6cd5\u3002<\/div>\n
\u968f\u7740\u4e8c\u4ee3\u6d4b\u5e8f\u4ef7\u683c\u7684\u964d\u4f4e\u4ee5\u53ca\u6355\u83b7\u65b9\u6cd5\u7684\u666e\u53ca\uff0cWGBS\u548cTarget-BS\u65b9\u6cd5\u8d8a\u6765\u8d8a\u5bb9\u6613\u5e94\u7528\u4e8e\u57fa\u7840\u548c\u4e34\u5e8a\u7814\u7a76\u3002<\/div>\n
\u76ee\u524d\u7684DNA\u5904\u7406\u65b9\u6848\u3001\u7532\u57fa\u5316\u6587\u5e93\u6784\u5efa\u65b9\u6848\u7684\u65b9\u6848\u8f83\u591a\uff0c\u6839\u636e\u4e0d\u540c\u7684\u6027\u80fd\u8981\u6c42\u548c\u5e94\u7528\u9700\u6c42\uff0c\u8981\u8bc4\u4f30\u548c\u9009\u62e9\u4e0d\u540c\u7684\u65b9\u6848\u3002<\/div>\n
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BS\u5904\u7406DNA\u7532\u57fa\u5316\u6d4b\u5e8f\u7684\u91d1\u6807\u51c6<\/div>\n
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\u5bf9DNA\u8fdb\u884c\u91cd\u4e9a\u786b\u9178\u6c22\u76d0\u5904\u7406\u662f\u6700\u5e38\u89c1\u7684DNA\u5904\u7406\u65b9\u5f0f\uff0c\u4e5f\u662fDNA\u7532\u57fa\u5316\u6d4b\u5e8f\u7684\u91d1\u6807\u51c6\u3002\u4f46\u8be5\u65b9\u6cd5\u6709\u5176\u660e\u663e\u7684\u7f3a\u9677\uff0c\u6bd4\u5982\u5904\u7406\u540e\u4f1a\u5bf9DNA\u9020\u6210\u65ad\u88c2\uff0c\u5360\u6bd4\u9ad8\u8fbe90%[2, 4, 5, 6, 7]\u3002<\/div>\n
<\/div>\n
\u4e3a\u4e86\u4fdd\u8bc1\u540e\u7eed\u6587\u5e93\u7684\u591a\u6837\u6027\uff0c\u5bf9DNA\u7684\u8d77\u59cb\u91cf\u6709\u8f83\u9ad8\u7684\u8981\u6c42\uff0c\u6240\u4ee5\u5728\u5904\u7406cfDNA\u6837\u672c\u8fd9\u79cd\u8d77\u59cb\u91cf\u8f83\u4f4e\u7684\u6837\u672c\u65f6\uff0cBS\u7684\u5904\u7406\u65b9\u6cd5\u76f8\u5bf9\u6bd4\u8f83\u56f0\u96be\uff1b\u518d\u6bd4\u5982BS\u5904\u7406\u540e\u7247\u6bb5\u65ad\u88c2\u6709\u5e8f\u5217\u7279\u5f02\u6027\uff0c\u4ee5\u53ca\u5bf9\u80de\u5627\u5576\u7684\u8f6c\u5316\u4e5f\u5e76\u975e\u662f100%\uff0c\u8fd9\u4e9b\u7f3a\u9677\u4e5f\u4f1a\u9020\u6210\u540e\u7eed\u7532\u57fa\u5316\u5206\u6790\u65f6\u7684\u504f\u5dee\u3002<\/div>\n
<\/div>\n
Nelly Olova\u7b49\u4eba[8]\u7814\u7a76\u4e86BS\u8fc7\u7a0b\u4e2d\u4e0d\u540c\u7684\u52a0\u70ed\u5904\u7406\u7684\u6e29\u5ea6\u3001\u65f6\u95f4\u3001\u53d8\u6027\u65b9\u6cd5\u7b49\u6761\u4ef6\u5bf9\u540e\u7eed\u6d4b\u5e8f\u7ed3\u679c\u7684\u8986\u76d6\u5ea6\u3001\u504f\u597d\u6027\u7684\u5f71\u54cd\uff08\u5982\u4e0b\u56fe\u6240\u793a\uff09\uff0c\u9ad8\u6e29\u52a0\u70ed\u548c\u9ad8\u6d53\u5ea6\u7684\u4e9a\u786b\u9178\u6c22\u76d0\u90fd\u4f1a\u5bf9DNA\u9020\u6210\u66f4\u4e25\u91cd\u7684\u635f\u4f24\u3002<\/div>\n
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\n\n
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\"RmFicvQBAkgBCnoXwL82fPGtN82mIqoKupFfWkAgibFFTd6s01icXLB5QibjvOYziaDjEd4DEYWia9jLLPwSLdNNPficA\"\/<\/figure><\/div>\n\n
\u56fe\uff1aBS\u8f6c\u6362\u534f\u8bae\u548c\u53c2\u6570<\/div>\n
\uff08\u6765\u6e90\/doi\uff1a10.1186\/s13059-018-1408-2\uff09<\/div>\n
\u6240\u4ee5BS\u548c\u4e8c\u4ee3\u6d4b\u5e8f\u7684\u7ed3\u5408\uff0c\u9700\u8981\u642d\u914d\u5408\u9002\u7684\u5b9e\u9a8c\u6d41\u7a0b\u624d\u80fd\u6709\u8f83\u4e3a\u7406\u60f3\u7684\u6570\u636e\u7ed3\u679c\uff0c\u5177\u4f53\u7684\u5b9e\u9a8c\u4e2dDNA BS\u5904\u7406\u7684\u524d\u540e\u987a\u5e8f\u5bf9\u7ed3\u679c\u4e5f\u662f\u6709\u5f71\u54cd\u7684\u3002<\/div>\n
Pre-BS(\u5148\u5efa\u5e93\u540e\u4e9a\u786b\u9178\u6c22\u76d0\u5904\u7406)\u7684\u610f\u601d\u662f\u5148\u5bf9DNA\u8fdb\u884c\u6253\u65ad\uff0c\u540e\u7eed\u8fde\u63a5\u63a5\u5934\uff0c\u540e\u7eed\u518d\u8fdb\u884cDNA\u5904\u7406\u3002\u00a0<\/div>\n
Post-BS(\u4e9a\u786b\u9178\u6c22\u76d0\u5904\u7406\u540e\u518d\u5efa\u5e93)\u7684\u610f\u601d\u662f\u5148\u8fdb\u884cDNA BS\u5904\u7406\uff0c\u540e\u7eed\u518d\u8fde\u63a5\u63a5\u5934\u3002<\/div>\n
Nelly Olova\u7b49\u4eba\u5728\u4e0d\u540c\u7684DNA\u5904\u7406\u65b9\u6848\u7684\u7814\u7a76\u57fa\u7840\u4e0a\uff0c\u4e5f\u7814\u7a76\u4e86\u4e0d\u540c\u7684\u6587\u5e93\u6784\u5efa\u6d41\u7a0b\u5bf9\u540e\u7eed\u6570\u636e\u7684\u5f71\u54cd\u3002<\/div>\n\n
\n
\"RmFicvQBAkgBCnoXwL82fPGtN82mIqoKuGKhFxFt7SmxmdH7BhoibmZOb1pUSiaSgMibP452ZAba3oJ2ELOQ0EnjsA\"\/<\/figure><\/div>\n\n

\u56fe\uff1aBS\u524d\u5904\u7406\u548c\u540e\u5904\u7406\u7684WGBS\u6587\u5e93\u5236\u5907\u65b9\u6848\u7684\u6570\u636e\u603b\u7ed3<\/p>\n

\uff08\u6765\u6e90\/doi\uff1a10.1186\/s13059-018-1408-2\uff09<\/p>\n

\u7814\u7a76\u4eba\u5458\u53d1\u73b0\u4e0d\u540c\u7684WGBS\u7684\u5b9e\u9a8c\u6d41\u7a0b\u4f1a\u6709\u975e\u5e38\u4e0d\u540c\u7684\u6570\u636e\u7ed3\u679c\uff0cpre-BS\u7684\u65b9\u6848\u6574\u4f53\u6570\u636e\u51fa\u73b0\u504f\u5dee\uff0c\u5c24\u5176\u662f\u5728DNA\u7684\u964d\u89e3\u4e0a\u3002\u8fd9\u548c\u6d41\u7a0b\u4e2d\u5148\u8fde\u63a5\u6587\u5e93\u63a5\u5934\uff0c\u518d\u8fdb\u884cDNA\u5904\u7406\u6709\u5173\uff0c\u8fd9\u6837\u4f1a\u6709\u5f88\u591a\u4e0d\u5b8c\u6574\u7684\u6587\u5e93\u65e0\u6cd5\u8fdb\u884c\u6d4b\u5e8f\u3002<\/div>\n
\u540c\u65f6\u4e5f\u53ef\u4ee5\u770b\u51fa\u5728Post-BS\u7684\u6d41\u7a0b\u4e2d\uff0camplification-free\u7684\u65b9\u6848\u4e0b\u4ea7\u51fa\u7684\u6570\u636e\u504f\u5dee\u548c\u51c6\u786e\u6027\u662f\u6700\u9ad8\u7684\u3002<\/div>\n
\u00a0<\/div>\n
\u5728\u5143\u7801\u57fa\u56e0\u76ee\u524d\u7684\u5e94\u7528\u4e2d\uff0c\u4f7f\u7528\u7684\u662fPost-BS\u65b9\u6848\u3002\u7531\u4e8e\u6837\u672c\u591a\u6765\u81ea\u4e8e\u4f53\u6db2\u7684cfDNA\uff0cDNA\u91cf\u8f83\u5c11\uff0c\u7ecf\u8fc7BS\u5904\u7406\u540e\u53c8\u4f1a\u5f62\u6210\u5927\u91cf\u65ad\u88c2\u7684\u5355\u94feDNA\uff0c\u9700\u8981\u5c3d\u53ef\u80fd\u7684\u63d0\u9ad8DNA\u7684\u6587\u5e93\u8f6c\u5316\u7387\uff0c\u6240\u4ee5\u5bf9\u4e0b\u6e38\u7684\u5efa\u5e93\u65b9\u6848\u8fdb\u884c\u4e86\u4f18\u5316\uff0c\u4f7f\u7528\u57fa\u4e8e\u5355\u94fe\u7684\u6587\u5e93\u6784\u5efa\u65b9\u6848\uff0c\u5bf9DNA\u5efa\u5e93\u8d77\u59cb\u91cf\u7684\u8981\u6c42\u5927\u5927\u51cf\u4f4e\uff08\u5982\u4e0b\u56fe\u6240\u793a\uff09\u3002<\/div>\n\n
\n
\"RmFicvQBAkgBCnoXwL82fPGtN82mIqoKuo3ahI2cJ2QbicLrxwuYXjzt3CzWHFia55vj6HaMibPQ1KjGOw7HMW3lbw\"\/<\/figure><\/div>\n\n<\/div>\n
\u56fe\uff1a\u5143\u7801\u6587\u5e93\u6784\u5efa\u65b9\u6848\uff08\u6765\u6e90\/\u5143\u7801\u57fa\u56e0\uff09<\/div>\n
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\u9176\u5904\u7406\u65b9\u6848\u66f4\u52a0\u6e29\u548c\u7684DNA\u5904\u7406\u65b9\u5f0f<\/div>\n
<\/div>\n
\u6839\u636e\u7814\u7a76\uff0c\u4f20\u7edf\u7684DNA\u7532\u57fa\u5316\u6570\u636e\u4f1a\u4ea7\u751f\u504f\u5dee\u7684\u6839\u672c\u539f\u56e0\u662fBS\u5904\u7406\u5bf9DNA\u6781\u5927\u7684\u635f\u4f24\uff0c\u6240\u4ee5\u66f4\u52a0\u6e29\u548c\u7684DNA\u5904\u7406\u65b9\u5f0f\u662f\u7814\u7a76\u7684\u65b9\u5411\u3002<\/div>\n
<\/div>\n
NEB\uff08New England Biolabs \u516c\u53f8\uff09\u63a8\u51fa\u4e86\u4e00\u6b3e\u4f7f\u7528\u9176\u8fdb\u884c\u5904\u7406\u7684\u8bd5\u5242\u76d2\uff0cNEBNext EM-seq\u5229\u7528\u9176\u4fc3\u8f6c\u5316\u6765\u53d6\u4ee3\u4e9a\u786b\u9178\u6c22\u76d0\u8f6c\u5316\uff0c\u80fd\u591f\u6700\u5927\u9650\u5ea6\u51cf\u5c11DNA\u635f\u4f24\u3002<\/div>\n
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\n\n
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\"RmFicvQBAkgAQ5R5PiaVYGPl2ALHHggPZNLyAxk64SlQ6DJeJg4dYFQG43pGyQM9OIiaDrON7cdjxicKsft2bopkMA\"\/<\/figure><\/div>\n\n
\u56fe\uff1a\u4e9a\u786b\u9178\u6c22\u94a0\u8f6c\u5316\u548cEM-seq\u6982\u8ff0<\/div>\n
\uff08\u6765\u6e90\/NEB\uff09<\/div>\n
EM-seq\u4e2d\u5bf9DNA\u7684\u9176\u5904\u7406\u5206\u4e3a\u4e24\u6b65\uff1a<\/div>\n
\u7b2c\u4e00\u6b65\u4f7f\u7528TET2\u9176\u4ee5\u53ca\u6c27\u5316\u589e\u5f3a\u5242\u5c065mC\u548c5hmC\u6c27\u5316\u62105caC\uff0c\u4fdd\u62a4\u7ecf\u8fc7\u4fee\u9970\u7684\u80de\u5627\u5576\uff1b<\/div>\n
\u7b2c\u4e8c\u6b65\u5229\u7528\u8131\u6c28\u9176APOBEC\u5bf9\u80de\u5627\u5576\u8fdb\u884c\u8131\u6c28\u5904\u7406\uff0c\u6b64\u6b65\u9aa4\u4e0d\u4f1a\u5bf95caC\u9020\u6210\u5f71\u54cd\u3002<\/div>\n
\u00a0<\/div>\n
\u6b64\u5916\uff0cEM-seq\u7684\u8f6c\u5316\u7ed3\u679c\u4e0e\u4e9a\u786b\u9178\u6c22\u76d0\u6d4b\u5e8f\u4e00\u81f4\uff0c\u6545\u53ef\u91c7\u7528\u539f\u5148\u7684\u6570\u636e\u5206\u6790\u5de5\u5177\u3002<\/div>\n
\u53e6\u5916\u4e00\u79cd\u9176\u5904\u7406DNA\u7684\u65b9\u6cd5\u662f\u725b\u6d25\u5927\u5b66\u8def\u5fb7\u7ef4\u683c\u764c\u75c7\u7814\u7a76\u6240\u5b8b\u6625\u5578\u548cBenjamin Schuster-Boeckler\u5f00\u53d1\u7684\u4e00\u79cd\u65b0\u7684DNA\u7532\u57fa\u5316\u6d4b\u5e8f\u65b9\u6cd5\uff1aTET\u8f85\u52a9\u5421\u5576\u787c\u70f7\u6d4b\u5e8f\uff08TET-assisted pyridine borane sequencing\uff0cTAPS\uff09\u3002<\/div>\n
\u00a0<\/div>\n
\u6b64\u65b9\u6cd5\u540c\u6837\u4e5f\u662f\u4e24\u6b65\u8f6c\u5316\uff1a<\/div>\n
\u7b2c\u4e00\u6b65\u4f7f\u7528TET\u9176\u5c065mC\u548c5hmC\u8f6c\u5316\u4e3a5caC\uff1b<\/div>\n
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[1].Frommer M, Mcdonald LE, Millar DS, Collist CM, Wattt F, Griggt GW, et al. A genomic sequencing protocol that yields a positive display of 5- methylcytosine residues in individual DNA strands. Proc Natl Acad Sci U S A. 1992;89:1827\u201331.<\/div>\n
[2].Grunau C, Clark SJ, Rosenthal A. Bisulfite genomic sequencing: systematic investigation of critical experimental parameters. Nucleic Acids Res. 2001;29:E65.\u00a0<\/div>\n
[3].Warnecke PM, Stirzaker C, Song J, Grunau C, Melki JR, Clark SJ. Identification and resolution of artifacts in bisulfite sequencing. Methods. 2002;27:101\u20137.<\/div>\n
[4].Raizis AM, Schmitt F, Jost JP. A bisulfite method of 5-methylcytosine mapping that minimizes template degradation. Anal Biochem. 1995;226(1):161\u20136.<\/div>\n
[5].Holmes EE, Jung M, Meller S, Leisse A, Sailer V, Zech J, et al. Performance evaluation of kits for bisulfite-conversion of DNA from tissues, cell lines, FFPE tissues, aspirates, lavages, effusions, plasma, serum, and urine. PLoS One. 2014;9:1\u201315.<\/div>\n
[6.]Shiraishi M, Hayatsu H. High-speed conversion of cytosine to uracil in bisulfite genomic sequencing analysis of DNA methylation. DNA Res. 2004;11:409\u201315.<\/div>\n
[7].Tanaka K, Okamoto A. Degradation of DNA by bisulfite treatment. Bioorg Med Chem Lett. 2007;17:1912\u20135.<\/div>\n
[8].Olova N , Krueger F , Andrews S , et al. Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data[J]. Genome Biology, 2018, 19(1):33.<\/div>\n
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