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Biology版 - another clone trick: 3-way ligation
相关主题
[合集] 郁闷,谁知道怎么回事Big fragment clonning
载体和fragment连接后的反应液能保存多长时间?Help! clone large fragment into pLenti7.3/V5 TOPO
ligase for large fragments问个克隆问题
How to check if my plasmid may or may not be table in DH5a?Big fragment cloning help?
怎样用infusion kit克隆多个PCR 片段??multisite gateway pro cloning kit from Invitrogen
Chip-seq DNA size 求助Synthesis of large DNA fragments
大片段连接求教Gibson assembly
4个片段连接的成功率有多少?help!dam methylation
相关话题的讨论汇总
话题: vector话题: clone话题: xhoi话题: ligate话题: ligation
进入Biology版参与讨论
1 (共1页)
l*****k
发帖数: 587
1
Well, I am sure I am not the first to do this, but I do want to share it
Sometimes, cloning multiple things into a vector can be tedious as you
have to transfer the fragments through several vector to get the
desired one, usually because the existence of the same Restriction site
in the vector and the fragment you want to clone.
say a gene has 2 frags in two separate vector, now you wanna get
them ligated into the same vector.
EcoRI------------XhoI Frag I (has interal BamHI site)
XhoI-
s******s
发帖数: 13035
2
I can tell u what I have done in China
Synthesis 10 ssDNA of 60Nts each (indeed, should be 5 dsDNA with
anealing end), ligate 6 together to get Mixture1, ligate the other
4 to get Mixture2, ligate Mixture1, Mixture2 with vector and finally
got the right product. It means to ligate 10 ssDNA (5 ds fragment)
with vector in one step, and it works!!!!
1 (共1页)
进入Biology版参与讨论
相关主题
help!dam methylation怎样用infusion kit克隆多个PCR 片段??
有什么RNA restriction enzymeChip-seq DNA size 求助
Re: Yeast Enolase_2 Help大片段连接求教
请教个蛋白间相互作用的问题,thanks a lot!4个片段连接的成功率有多少?
[合集] 郁闷,谁知道怎么回事Big fragment clonning
载体和fragment连接后的反应液能保存多长时间?Help! clone large fragment into pLenti7.3/V5 TOPO
ligase for large fragments问个克隆问题
How to check if my plasmid may or may not be table in DH5a?Big fragment cloning help?
相关话题的讨论汇总
话题: vector话题: clone话题: xhoi话题: ligate话题: ligation