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microsatellite loci amplification

kfeldh1 at ICARUS.CC.UIC.EDU kfeldh1 at ICARUS.CC.UIC.EDU
Wed Feb 26 15:45:55 EST 1997


I am trying to amplify lemon shark microsatellite loci using the PCR but 
haven't had much luck.  The DNA I am using was extracted from fin clips 
(stored in ethanol), by standard phenol-chloform extractions.  The 
primers being used are specific for lemon sharks (were developed from a 
lemon shark gDNA library).  

We are getting plenty of DNA from the extractions, so I don't think lack 
of DNA is a problem.   

I have tried many different PCR "recipes" (using DMSO, spermidine, 
varying molarity of dNTPs, varying volumes of PCR reaction, etc.) with 
no luck.  Does anyone have any suggestions?  Could something be 
co-amplifying with the DNA that is inhibiting the PCR?

Thank you in advance,

Kevin Feldheim



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