p**h
发帖数: 99 | 1
that is a tricky stuff.
there are many methods for site-directed mutagenesis. what is in your kit?
anyway,
make sure your oligos are correct (like it can not form a hairpin structure,
ect.)
if possible, you may try other kits. my favorite was a kit from promega
(forget its name), never lost with it. hehe. I hate PCR method (some ones
like it). | o******t
发帖数: 68 | 2 1.别人用的是同样公司出的kit吗？
2.你的模板有没有问题？反应体系好好检查。
3.引物是否合理？ | a****a
发帖数: 3905 | 3 换个kit吧, promega的很好用. 你是为了mutation又不是为了研究它的kit,
没必要在这上面化时间. | y***n
发帖数: 99 | 4 Why not do as someone said before,
synthesis two oligo with mutated nucleotide, | s**********i
发帖数: 711 | 5
quickchange doesn't use PCR actually. the product is not exponentially
to cycles numbers.
make sure that your primers have high enough Tm, (80C). do all the controls.
try incresing extension temp to 72C. make sure the extension time is long
enough. |
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