b******r
发帖数: 111 | 1 Hi,everyone,
I have to use single restriction sites for insertion of multiple fragments
. When using KpnI site, just get one successful insertion from 70 clones.
For NotI, it is 1 from 4,BamHI is 1 from 2. My question is why the ligation
is so bad at KpnI site? Or other reasons? Thanks | b******s
发帖数: 1089 | 2 实验室其他人遇到过经年的KpnI不work。但我自己切没遇到过问题。单酶切你可以上很
浓的质粒，多加点酶，但是不要超过反应体系的10%，过夜，加CIP，应该能做出来。
fragments
ligation
【在 b******r 的大作中提到】
: Hi,everyone,
: I have to use single restriction sites for insertion of multiple fragments
: . When using KpnI site, just get one successful insertion from 70 clones.
: For NotI, it is 1 from 4,BamHI is 1 from 2. My question is why the ligation
: is so bad at KpnI site? Or other reasons? Thanks
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