cDNA library (in Lambda ZAP vector) screening - picking up plaques on nitro cellulose membrane as replica (Dec/30/2001 )
while taking replica (second lift)of labda phage (cDNA library) on a nitrocellulse membrane for screeing by antibody the soft agar comes out with the membrane .if there is any solution to avoid this?
Looks like a rather simple answer, but sometimes these answers are the best. Precool your plates for about an hour before using them. I experienced the same problems in my lambda plaque lifts and by cooling the plates the problems vanished!
Hope that I helped you
Sometimes precool the plate is not enough to prevent the top agar from adhering to the membrane, for this reason, I usually add more NZY top agar to the LB/NZY plate than the Kit mentioned when spread the plaques.
Hope this help you!