Chemicon公司的CpGenomeDNAModificationKit的protocol修正
我用了快4个月这个试剂盒了，下面是一些总结出来的经验，我现在诱变每次都很成功了，呵呵：）1. 我溶NaOH的水PH值大概在6.5左右，所以我每次调Reagent I的时候3MNaO0H基本都要加150uL左右。2. 解链时用50度水浴15分钟。3. 加入Reagent I后，50度水浴16小时。4. 加入Reagent II和III后，室温孵育10分钟5. 第一次离心用9000 rpm 5分钟。6. 70%乙醇清洗3次，每次9000rpm 1 分钟，最后一次13000rpm2min。7. 加入20mM NaOH/90%EtOH后，反映5分钟。8. 不要去掉20mM NaOH/90%EtOH，直接加入90%乙醇后离心，9000rpm 1min9. 90%乙醇清洗2次，每次9000 rpm 1 min，最后一次13000rpm 3min10. PH 7.5 TE buffer 65度 溶解 15 minPS: 如果发现ＰＣＲ结果中M......阅读全文
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