3 replies to this topic

[Wek]

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

[bob1]

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

[Wek]

Is there any protocol to optimize the DNA for sequencing?

[phage434]

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.