How do I PCR a DNA fragment with >200 CGG repeats? - (Jan/15/2011 )

Hi there,

I'm trying to amplify the triplet repeat region of a Fragile X individual who has >200 (CGG) repeats. This seems to be problematic, so any help with regards to primer design and PCR cycling parameters or PCR conditions would be much appreciated. What modifications would I need to make to a standard PCR reaction, and if possible, why would I make them... this would help me understand the rationale behind it all. I hope someone is able to help me figure this out.

Many thanks

2bornot2b on Sun Jan 16 00:30:58 2011 said:

Longer denaturation times in the cycles, some additives (DMSO, betaine (it is supposed that this is the main ingredient of the "Q-solution" from qiagen or "GC-RICH solution enhancer" from Roche)).