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primer labelling(5' end) and hybridization
Posted by: chinmay (IP Hidden, New member, 2)
Date: January 19, 2006 03:38AM

hi everybody
i am looking for a sequence(attRS1) that is essential for integration of ctx phage in V. cholerae genome in some of my strains. the attRS1 sequence is 18 nucleotide sequence, i have that nucleotide sequence as a primer. so what i am trying to do is 5' end labelling (gamma 32 P using PNK) that fragment and using as a probe in southern hybridization. but i m not getting the signal, either i m getting a dark background or complete blank. couldn't really understand where the problem is, checked everything, but didn't got a clue. checked with PNK from promega and USB. primer is intact(checked).
bye

 

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