T4 (without BSA) - How efficient the T4 is? (Sep/27/2007 )
I was doing clonning without the BSA in my T4 reaction mix.
Anybody have nay idea?
Thanks in advance!!!
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.
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.