IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

homemade T addition or blut PCR cloning?

Shin Enomoto shin at biosci.cbs.umn.edu
Sat Dec 2 17:00:14 EST 1995


I want to clone a PCR fragment into a expression vector I made.  The 
polylinker has a Pvu II site and need to make a in-frame fusion into this 
site.  
1) Clone the PCR fragment as blunt by using a polymerase that does not 
give over-hangs
2) Clone the PCR fragment by first polishing the extra base and blunt clone
3) Add a extra T to the pvu II site either by taq polymerase or terminal 
deoxy transferase and clone in a PCR fragment with an A over-hang.
4) Design a restriction site into the PCR primers to clone

Which one of the above is likely to be the easiest? I will summarize the 
responses.

Thank you




More information about the Methods mailing list

Send comments to us at biosci-help [At] net.bio.net