8楼: Originally posted by gyesang at 2013-12-09 00:59:54
有载体自连，那你酶切载体的量是多少呢？
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5楼: Originally posted by zh10246 at 2013-12-06 15:13:04
YOU CAN DO sequential digest.

12楼: Originally posted by yanzixiang at 2013-12-09 10:34:47
能说的具体一些吗，不太懂...

15楼: Originally posted by yanzixiang at 2013-12-09 06:36:03
比如我先用bam H1切一段时间，切开后Xba 1那个酶切位点上只有一对G\C可以看做是其保护碱基，但是当G\C作为xba1的保护碱基时，酶切2小时切开率也还是0%（参考NEB保护碱基表），这样能切开吗？先切Xba1也是这样。

16楼: Originally posted by b6122 at 2013-12-10 02:41:55
那么能不能重新构建载体，在当前位点上做site directed mutation，插入更多的保护碱基
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17楼: Originally posted by yanzixiang at 2013-12-09 21:07:12
不知道怎么操作...