WGA kits - experiences? (Jan/12/2010 )

We want to

1) amplify genomic DNA in order to get more mitochondrial DNA starting material for LongRange-PCRs and cloning/Sanger-sequencing
2) amplify ds cDNA samples for 454 sequencing

using a whole genome amplification kit. Does anybody has experience in this field and can recommend a WGA-kit? Qiagen, Sigma, GE Healthcare, ...?

Concerning the mt-DNA: we have very small and few animals as starting material. Does anybody know if the REPLI-g Mitochondrial DNA kit (Qiagen) works for non-human material, too?

Thanks in advance!

-confused-

confused on Jan 12 2010, 01:36 PM said:

Hi
We use WGA kit from Qiagen.
We obtain up to 5 microgram DNA from 0.1ng human genomic DNA.
DNA quality seems to be good on gel.
However, Nanodrop measurement is misleading.
Most probably because of primers remaining in the reaction even after ethanol precipitation.
We are now trying to use ExoSap to clean the reaction before ethanol precipitation.
Hope it is helpful
Best
Michael

-Michaelro-

I've used the Sigma WGA kit and once compared it with the ligation mediated PCR of gDNA. comparison was done by a whole genome microarray with the WGA and LMPCR of the same DNA. the results were pretty comparable for both and we moved from LMPCR to WGA as it required much less DNA and much easier to perform. the yields were approx 7.5 ug from 50 ng.

-gogreen-

Hi,

Perhaps you can consider GE Healthcare Genomephi Hy DNA amplification kit.

-stylothecancer-