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PCR from protozoa DNA


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#1 nhiphops

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Posted 17 February 2014 - 11:55 PM

This for my first time to amplification dna from protozoa (human dna, i'm ok). I found any problem about low yield of dna extraction from stool, very hard to amplification.

i would like to talking about that and want to share your experience.

 

Thank you



#2 Trof

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Posted 18 February 2014 - 10:45 AM

Stool contains lots of PCR inhibitors. You may try adding BSA (bovine serum albumin) to the sample (10ug/20ul reaction) to block them nonspecifically.

 

Also some species can have very different genomic GC content, than what's usual for mammalian DNA. Very low or high GC has different solutions. You can check your amplicon and find out.

 

You may also search on forums for protocols on DNA isolation from stool.


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#3 hobglobin

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Posted 18 February 2014 - 12:46 PM

The question is also how much protozoan DNA you can get since protozoa content might be very low and variable...most DNA will be bacterial and human (if it's from humans).


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#4 nhiphops

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Posted 19 February 2014 - 07:00 PM

Thank you for good answer. I'll try to do it again.

(Our Stool samples're from humans.)






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