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-   -   Problems in screening genomic DNA Library with antibody (http://www.molecularstation.com/forum/molecular-cloning-forum/72254-problems-screening-genomic-dna-library-antibody.html)

srivels 01-17-2010 04:42 PM

Problems in screening genomic DNA Library with antibody
 
I constructed a genomic DNA library of a bacterium using Partial DNA digestion employing Lambda ZAP II Kit (Stratagene). I did everything alright as shown in kit. I used 2-4 kb fragments for ligation with pre-digested vector supplied in kit. I used 1 uL of ligate for packaging and then amplified the library. In Blue-white screening, all the plaques showed transperant or clear with blue rings at the edges. There was no blue colony. I thought all are recombinant clones. Then, I started screening the library using antibodies. But I am not getting any positive clones. Please any one help me and tell the reason? My antibodies are good and I checked them several times.:headbonk:


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