MadSci Network: Molecular Biology

Re: how do you design primers for PCR if you don't know what the DNA sequence?

Date: Mon Mar 3 01:39:52 2008
Posted By: Sebastien De Landtsheer, Undergraduate, Immunology, Laboratoire National de Sant�
Area of science: Molecular Biology
ID: 1204419571.Mb


It's quite simple : if you do not have the actual sequence of the DNA you 
want to amplify, or at least the 3' and 5' ends, then you cannot design 
primers and run conventional PCR.
But there are many ways to amplify DNA. Search your literature for the 
following keywords : SISPA, phi29 polymerase, multiple displacement, RDA, 
and isothermal amplification. I advise you the papers from Allander et 
al., which are quite detailed and give a good explanation...


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