f********g
发帖数: 6 | 1 You don't need kinase if the vector/fragment the linker is ligated to has 5'p
on its end, which usually is the case. When you use restriction enzyme to
generate the new end from the linker, you also generate the 5'p on the end. I
think people who are using kinase are ligating oligos to de-phosphorylated
vector etc, in that case you need to add 5'p to your annealed oligos.
我以前anneal以后直接用来做连接，从来没有问题。而且anneal过程中我没有加过kinase | l*****i
发帖数: 429 | 2 我就用水溶解，连起来也没问题。
２５uM working concentration for each one, boil at 100 degree for 5 minutes,
let it cool naturally.
Tried at least 5 times. |
|
