rbcL PCR in cyanobacteria

p.ravenscroft p.ravenscroft at clara.net
Wed Apr 15 15:54:10 EST 1998


on 15 Apr 98, Ashbwill wrote...

>If anyone has had any success in PCR amplification of rbcL gene in
>cyanobacteria, please help me.   I am having a great deal of difficulty getting
>priming.  Primers look fine, DNA looks fine... Something is up.  Any advice?
>
Never tried it with the gene in question, however I had some problems with
PCR's, so here goes.

Have you tried altering perameters such as temperature of primer annealing. I
had great difficulty with such a problem. First tried "Touchdown PCR", altering
number of cycles at different temperatures, until I could identify a good
annealing temperature.

Could also try altering MgCl concentrations




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