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T4 (without BSA) - How efficient the T4 is? (Sep/27/2007 )

Hi,
I was doing clonning without the BSA in my T4 reaction mix.
Anybody have nay idea?
Thanks in advance!!! ph34r.gif

-Niraj-

Nope no idea. I've never made a comparison. I don't have BSA in my T4 ligase buffer. I would guess the BSA reduce the amount of T4 ligase protein adsorbed by the plastic walls. Though, T4 ligase is sold at quite high concentrations.

Hmm...

-perneseblue-

L. F. Peruski, Jr. reports that BSA stimulates ligation for unknown reasons. (L. F. Peruski, Jr. 1992 USB Comments 19 (1) 4-6.)

In my hands, with limited amounts of vector and insert, adding BSA increases the number of colonies obtained, probably because it prevents the absorption of DNA on the tube walls.

-tfitzwater-