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Best way to genotpy a mutant gene with a 1bp difference. - (Jul/20/2012 )

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.



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


Is there any protocol to optimize the DNA for sequencing?


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.