cDNA library--questions

Chi-kuang Wen ckwen at expert.cc.purdue.edu
Fri Oct 18 13:04:26 EST 1996


 Dear Netters:

Currently I have a problem with my cDNA library, and any suggestions would be
appreciated.

The size of the cDNA library is fine,but the only thing bothers me is smalle
size of the plaques (<1mm).  The positive control (wt lambda) was included and
large plaques were obtained (>2mm). It, therefore, seems that my media are
fine.

I was told that using LB(-yeast ext) top agar may help.  It did increase the
size of the plaques, but the plaques become cloudy, which means that the 
phage number per plaque may not be increased although the size of the plaques
is bigger.

Prolonged culture did not increase the size of the plaques.(>24hr at 37C).

The vector I used is lambda ZipLox RI arms, and the E.coli strains are
either LE392 or Y1090(ZL). BOth NZY and LB plates have been tried.
The cDNA inserts are 0.5 to 1.6Kb, by PCR several plaques.

Thanks for any help,

Chi-Kuang
.
 



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