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How to joint two cloning genes?Moderator: BioTeam
4 posts • Page 1 of 1
How to joint two cloning genes?I want to construct a fusion gene, which comes from chromosome translocation.
I have the sequences of the two DNA fragment. But how can I joint them together? One ends AACAGG One begins TCCTCC I input the sequence AACAGGTCCTCC, and try to find a restriction enzyme to cut them. But they gave me four enzymes: AvaII, EcoO109I, PpuMI, Sau96I. The sequence of Sau96I is GGNCC. What does N mean? Can I use this enzyme to joint two genes? Or is there any other way to joint the two genes? I don't want to add enzyme site between two genes, for that maybe interfere the fusion gene's function.
Re: How to joint two cloning genes?Blunt Ended Ligation?
You could try Slicing by Overlapping PCR (SOE-ing), its very easy. You just need one overlapping primer
Source: Karin L Heckman & Larry R Pease. Gene splicing and mutagenesis by PCR-driven overlap extension. Nature Protocols 2, - 924 - 932 (2007)
4 posts • Page 1 of 1
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