发信人: sichou (nini), 信区: Biology
标 题: Re: help!a question on cloning!
发信站: BBS 未名空间站 (Mon Nov 14 00:53:34 2005), 转信

i've been handling alot blunt end ligation this semester. several advice mig
ht be helpful to u:
1. use 1:10 or even higher ratio
2. for me, i use roche rapid ligation kit it works fine so far
3. mix the vector and blunt-end fragment @ RT for 1h or longer (O/N) IN THE
PRESENCE OF THE RESTRICTION ENZYME U USE TO CUT THE VECTOR, which will drama
tically kills most of the auto ligation.


【 在 apple12 (apple) 的大作中提到: 】
: I wann clone a 16Kb blunt ends fragment. I need some ideas and advices from
da
: niu!
: Thx!



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