Vent Polymerase

Zhonglin Chai zchai at COBRA.PATH.MONASH.EDU.AU
Sat May 31 21:21:57 EST 1997


>Dear All,
>I wonder if you could help me in this point. When I want to remove a
>cohesive end froms my plasmid by using Vent Pol.,should I add dNTPs or
>not?
>Thanks

Yes, you do. I think Vent and other enzymes with proof reading activities
have both 5' -> 3' polymerization activity and 3' -> 5' exonuclease
activity. As soon as the 3' -> 5' exonuclease activity delete the nt from
the 3' end and leave the opposite DNA strand with a 5' overhang, the 5' ->
3' polymerization activity would add nt to the 3' end according to the
opposite strand sequence, that needs dNTPs as building materials.



Dr. ZhongLin Chai
____________________________________________
Department of Pathology and Immunology
Monash University Medical School
Alfred Hospital
Commercial Rd, Prahran, VIC 3181, AUSTRALIA
Telephone: (61 3) 9276 2698 (lab)
           (61 3) 9276 2696 (office)
           (61 3) 9384 1305 (home)
           0419 87 1940 (mobile)
           + 61 419 87 1940 (international mobile)
Fax:       (61 3) 9529 6484
email:     zchai at cobra.path.monash.edu.au
____________________________________________





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