DNA extraction for NGS from small samples

[samcd]

Hi

Has anyone attempted DNA extraction on small samples (say 500-1000 cells) for any NGS protocol?

We currently attempting to extract DNA from single human colonic crypts and for our targetted sanger sequencing its fine to use picopure but is it ok, or are there better methods? Most column based kits yield no to little DNA.

Any advice would be welcomed

[Calluna]

I've used the NEB Single Cell WGA kit to extract and amp <10 cells. It's expensive, but seems to work well.

https://www.neb.com/products/e2620-s...kit#tabselect0

[NextGenSeq]

We've done RNA-Seq from 50 cells from LCM samples. Fluidigm cells a C1 autoprep for single cell RNA-Seq using the Clontech SMARTer kit. For DNA-Seq check out the Diagenode Microplex kit.

[samcd]

Thank you both.