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Old 07-17-2013, 06:33 AM   #1
samcd
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Location: london

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Default DNA extraction for NGS from small samples

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
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Old 07-17-2013, 07:09 AM   #2
Calluna
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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

Last edited by Calluna; 07-17-2013 at 07:12 AM.
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Old 07-17-2013, 07:38 AM   #3
NextGenSeq
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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.
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Old 07-18-2013, 04:18 AM   #4
samcd
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Thank you both.
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