I would like to know if Psychrophiles obtained from environments ranging from 0-10 degrees contain more of GC in their genome. I have community DNA (microbial) from cold adapted regions but the conventional PCR fails to give me any amplification. Is there any co-relation? How do I find out if I have to opt for PCR protocols for GC rich templates?
Thank you.
In my experience there is no direct correlation between psychrophiles and GC content. However it is possible that you have something like an Arthrobacter species, some of which are phychrophiles and known to have high G+C content.
About your PCRs, trying running them with a 5 minute denaturing step (94 or 95C) before going into your cycling steps. This is assuming that your primers, reagents, etc. are ok. Check out the following paper about isolation/identification of psychrophiles: http://aem.asm.org/content/70/1/202.full?sid=820e82a4-ff1b-4f1e-9723-bde1dd5c0aa8. It has a good PCR protocol and several primers for you to try.
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