Lambda EMBL 3 primers

Alberto Carboni carboni at imiucca.csi.unimi.it
Tue Feb 27 03:10:02 EST 1996


Hello netters,

does anyone experienced PCR amplification of inserts
in a genomic library in lambda EMBL 3 vector?

If so can someone send me the sequence of PCR primers 
designed on the R-arm and L-arm of the vector?

The problem raises from the discovery that for excising
the genomic insert from the recombinant vectors I must use
the RE SalI, wich is sensitive to PEG contamination. In pre-
sence of small traces of PEG, SalI gains the so called 'star
activity', and recognises BamHI and EcoRI sites.

Thats why I obtain, form a single positive colony, a ladder of
restriction fragments. 

I decided to PCR amplify my insert and than clone it in a
plasmid vector.

Any suggestion would be appreciated.

Thank you,

Alberto




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