Question about DNA extraction and PCR

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Question about DNA extraction and PCR

Postby nanff » Oct 10 2017 9:13 pm

Hi I am a new one in DNA extraction and PCR.

I extracted DNA from mouse tail which should contain one control Genome and one (or two) of two known target Genomes.
After PCR, the control genome always showed in Electrophoresis which indicate the exsitance of the chromosome. However, none of the target genomes showed in the electrophoresis result.
I also did sample of the two target genomes with the same primers I used and both showed in electrophoresis result. So it should not be primier issue either.

Can anyone help me check anyother posibility of not-showup of the target genomes?
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Re: Question about DNA extraction and PCR

Postby r.rosati » Oct 11 2017 8:56 am

That's a bit vague. Have you ruled out the possibility of spurious priming sites on "genome 1" for the primers amplifying genomes "2" and "3"?
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Re: Question about DNA extraction and PCR

Postby nanff » Oct 11 2017 3:07 pm

r.rosati wrote:That's a bit vague. Have you ruled out the possibility of spurious priming sites on "genome 1" for the primers amplifying genomes "2" and "3"?

Thanks for your comment. I did PCR for genomes 1, 2 ,3 respectively, and all the primers works well in previous sample.
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Re: Question about DNA extraction and PCR

Postby 29yrsExperience » Oct 12 2017 12:16 pm

Hi! Is there anything about this particular mouse that is different from the other mice you have tested? A different strain, perhaps? Are you sure the primers bind in places that are conserved in all strains? We could help you more if we knew more about your gene targets and what your experiment is designed to test. How large are the amplified fragments you expect to see, for instance?
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