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!!!! |
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