How much DNA per plaque?

stephane leclair at mpiz-koeln.mpg.de
Tue Sep 23 03:08:58 EST 1997


Hi, all,

I am doing plaque lifts from a lambda cDNA library. The library has been
produced with a Stratagene kit and uses the "Uni ZAP XR" vector (phage).
Recently, I started to have doubts wether my probe is good enough to
detect DNA amounts that are common in plaque lifts. Increasing the
concentration of the probe during hybridization or prolonging the
hybridization time does not help.

How much DNA per plaque binds in average on a nylon membrane?

I am aware that this depends (among other factors) on the lay-on time of
the membrane but I am interested in a crude number only. My guess is
something in the lower nanogram range.

Any comments are welcome.

Thanks						Frank



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