发信人: leohawk (leohawk), 信区: Biology
标 题: another clone trick: 3-way ligation
发信站: The unknown SPACE (Fri May 3 13:38:44 2002), 转信

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-------------BamHI Frag II(has internal EcoRI site).

just cut a vector with EcoRI/BamHI and put all three fragment in the
ligation system...

you know what, it works with amazing efficiency! My boss was very amazed
when he see it worked so efficiently.

never tried 4-way clone, but it might worth a try....

hehe...

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