Inhibitor in the PCR reaction?

Khursheed Sohrab Gimi ksgimi at whale.st.usm.edu
Thu Oct 10 11:33:21 EST 1996


Summary:Can someone help with this problem we are having with our PCR
reaction? We are trying to develop a method to detect viral infection in
Wild Penaeus aztecus shrimp. However, although our controls work and
although the same PCR procedure works for lab reared shrimp, it does not
for the wild shrimp. Both are infected with the same virus. It may be an
inhibitor problem and we have tried several DNA isolation procedures:
Boiling with chelex, proteinase K digestion followed by chl/ph extraction
followed by ehanol precipitation, even a commercial prepration (Isocode
stix used for blood) and several others.  However, the problem still
persists so we tried modifications to the PCR like heat soaked PCR,
Hrdrogen peroxide treatment (used for blood), addition of BSA and addition
of DMSO and glycerol among others, to the reaction mixture. If anyone has any
suggestions, I'd appreciate it. 
	Thanks.
  


 
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-- 
Khursheed Gimi
University of Southern Mississippi
Dept. of Biological Sciences
Hattiesburg, MS.
Internet:  ksgimi at whale.st.usm.edu



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