hi Ariananda,

it should not be necessary to have a single product  as you optimise the running conditions to analyse the fragment that you are interested in but it is easier with a unique product.I will have a look for information but ususlly an email to the company selling the equipment (transgenomic wave in my depts case) will always get useful information

bw

paul

Very thankful for your answer, Paul

I am trying to detect NPM1 and CEBPA mutation with DHPLC. I have settled the method to detect NPM1 mutation, but it is quite tricky for CEBPA. Because the sequence is quite long, GC-rich and I have to divide it into 3 separate region. I am using a method that has been optimized and published in Blood. But still, it is quite hard to reproduce the method.

I really need some reference to use DHPLC for mutation detection, since I never have any. I will try to contact transgenomic to get more information.

Thanks Paul,

Regards,

Aria

Hi Aria,

cebpa has 4 polymorphisms...can you use any of these to show that the technique is working. It is very gc rich which is a problem but when I was doing denaturing gradient gel electrophoresis one method of moving a melting domain into the area of interest was to add a gc tail to one of the primers which binds one end of the product and forces the other end to melt preferentially.Possibly a similar methodology might work here but your sequence is very gc rich so will be quite difficult to melt.

good luck

paul