PCR in plant genomic DNA

Philip E Dykema pedykema at iastate.edu
Fri Sep 3 16:19:06 EST 1993


___________________________
 Does anyone has a protocol for PCR in plant genomic DNA ? We have some
problems. Our PCR control worked but the PCR in plant genomic DNA didn't work.
                                                 Thank you
                                                  Parin
__________________________

Well, I guess its time I introduce myself as well.  My project involves 
investigating potential homologs to an embryogenic gene (LEA, Group I to be 
specific).  I am using both Southern blots and PCR analysis on a host of
plants from all of the divisions in Kingdom Plantae.

As for your question, Parin - I have found that the PCR results depend 
primarily on the primers used.  Are you using primers designed for one specie
only, or are the primers "degenerate" (that could stir some debate among
the free will discussion!) ?  Have you tried different annealing temperatures
in separate reactions; I have just obtained some evidence for my own primers 
showing differing band patterns at different temperatures.  Also, what was the
control template - plasmid, insert, genomic clone?  I guess I need some more
information.

Philip Dykema (pedykema at iastate.edu)
Iowa State University
Botany Department



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