Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.
Fri Feb 02, 2007 7:50 am
We have a patient with a silent mutation, but the sequence change does not create/abolish any restriction enzyme site. We tried allele specific PCR, to check if the silent mutation is present in the normal population, but my student is having a hard time with the optimisation. Its not that she's doing allele specific PCR for the first time - she did it for so many other genes. She also tried hot start PCR but failed to optimise it many times.
We dont think buying "magic PCR premix" would be something promising. Anybody has any idea any other methods to screen the normal population for such sequence change? TIA.
Tue Feb 20, 2007 2:46 pm
SSCP (Single-Strand Conformation Polymorphism try it, it may work for you
Wed Feb 21, 2007 4:15 am
I guess from what you say that your mutation do not change the length of the product either, just the sequence. In this case DGGE/TGGE could work. But The problem is that it might require some investment to start. It might be worth investigating though.