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Best way to genotpy a mutant gene with a 1bp difference.


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3 replies to this topic

#1 Wek

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Posted 19 July 2012 - 09:20 PM

What is the best way to genotype a mutant gene with a 1bp difference? I'm trying to check whether my DNA is homozygous or heterozygous mutant but I'm having some difficulties due to the tiny difference.

Thanks

#2 bob1

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Posted 20 July 2012 - 12:02 AM

You could either do sequencing (most sensitive) or perhaps dHPLC if you have one available.

#3 Wek

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Posted 20 July 2012 - 05:18 PM

Is there any protocol to optimize the DNA for sequencing?

#4 phage434

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Posted 21 July 2012 - 01:58 PM

You may be able to RE cut the fragment to make it short, e.g. 25 bp. The 1 bp difference would then be far easier to see on a high concentration page gel.




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