Bovine Serum Albumin in PCR

Dr. Duncan Clark duncan at genesys.demon.co.uk
Sat Nov 9 06:04:06 EST 1996


In article <19961106204600.PAA23917 at ladder01.news.aol.com>,
jenova at aol.com writes
>
>Can anyone tell me what the purpose of bovine serum albumin in the PCR 
>reaction could be? This reagent is listed in published paper on genotyping
>of Clostridium difficile. Also, how much of this reagent should I add to 
>the PCR reaction mixture? 
>


Two possible reasons. 

2.5u of Taq pol is 0.01ug of protein so addition of BSA
increases the protein concn.

Addition of BSA reduces non-specific binding of the enzyme to plastics,
glass etc. by coating those surfaces itself. 

How much to try? Idaho suggest using 2.5mg/ml BSA in a 10x buffer for
their hot-air machine so start with that. Do not use acetylated BSA just
heat treated for say 80C for 20mins (you are just trying to heat kill
DNAse and protease etc.) Some BSA's will ppt when doing this so try a
few sources. Acetylated BSA will kill your polymerase.

Duncan


--------------------------------------------------------------------------------
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Dr. Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.prestel.co.uk
http://www.genesys.demon.co.uk



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