Whole Genome Amplification

I used Whole Genome Amplification (WGA) to recover some very old DNA and include a maximum of samples into GBS plates for a mapping project. I will do another post about that, but here I would like to resume how I did the whole genome amplification.

For information on how the whole genome amplification method works, check the previous post by Moira.

I used the Qiagen Repli-g Mini Kits with the amounts divided by half.

The protocol is the following:

2.5 ul of DNA

2.5ul of buffer D1 (DLB + nuclease-free H2O)

Incubate for 3min at room temperature

Add 5ul of buffer N1 (stop solution + nuclease-free H2O)

Vortex and centrifuge

Add 15ul of master mix (repli-f reaction buffer + DNA polymerase)

Incubate at 30oC for up to 16h and inactivate enzymes at 65oC

I found that 6h was good for the amount of DNA I was hoping to recover. I recovered at least 50x the amount I started with, typically starting with concentrations between 1-10ng/ul and getting on average 60ng/ul out of the WGA protocol.

Depending on the use of the DNA, you may want to play with the incubation time (e.g. keep it short), especially if you plan to use PCR based approach later on (almost everything is PCR-based..).