5 replies to this topic

[scbody]

Hi
I'd be interested in people's experiences of commercial kits for genomic DNA isolation from human whole blood (5-10 mL) or buffy coat (~500uL); especially cost, time and reliability issues.
Thank you

[wulinteo]

Qiagen kit .. is the best to extract Human DNA

[scbody]

Thank you

[FionaGould]

I used Qiagen, though if you need to screen for PCR, you can add 900ul 1xSSC to 100ul blood, mix then pellet by centrifugation. Decant supernatant, add 100ul of a lysis buffer and 15ul Proteinase K. Incubate O/N at 60oC, vortex well, pellet as before and put 1ul of supernatant into 49ul water. This gives approx 5ng/ul. Incubate at 95oC 5 mins to deactivate PK. This is good enough for PCR.

[karen pooley]

Hi
Nucleon kits are excellent for DNA extraction from 0.5ml to 10ml of whole blood

kap28@mole.bio.cam.ac.uk

[scbody]

Karen and Fiona
Thank you for your input.
Regards
Simon