发信人: adeno (天夺其魄), 信区: Biology
标 题: Re: 有没有做4-piece ligation的?
发信站: Unknown Space - 未名空间 (Wed Nov 12 23:47:47 2003) WWW-POST

Nothing special. Inserts ratio should be 1:1:1. Vector amount around ~50ng.
Inserts the more the better.

I use epicentre's fast-link ligase. I do three-way ligation routinely and
never failed once. Once for a while I do 4-way ligation, sometimes even with a
blunt end:)

【 在 qiangguo (fuming) 的大作中提到: 】
: any tips? thanks.
: 【 在 windywendy (山田樱子) 的大作中提到: 】
: : I just did a 5-piece ligation, got 1 positive among 10. Worth of trying.
: : 【 在 kiddingyou (李寻欢没刀了) 的大作中提到: 】
: : : one step? good luck. My last 3 piece almost killed me. Maybe I was
stupid.
:
:

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