PCR not working!! !!!

bic_nii at imtech.ernet.in bic_nii at imtech.ernet.in
Tue Oct 26 13:32:54 EST 1993

Dear netters,

I have been facing problems in getting consistent PCR results. I have
3 sets of primers derived from partial sequencing of a recombinant 
clone. I'm trying to amplify a small region of this clone using 
different permutations of the primer sets. I have tried varying 
concentrations of Magnesium, primers, DNA templates, different 
temperature range and even different PCR machines, but the results 
show the same inconsistancy. Sometimes the amplification occurs in the 
expected range and sometimes it doesn't work at all. One problem could 
be non-specificity of the primers but I have got good sequencing 
results with the same lot.  
This is an SOS for help. Any suggestions are welcome.
Thanx in anitcipation.

gauri at nii.ernet.in
National Institute of Immunology,
Aruna Asif Ali Marg,
New Delhi-110 067,

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