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Human whole blood genomic DNA extraction kit


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5 replies to this topic

#1 scbody

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Posted 28 May 2002 - 12:33 PM

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

#2 wulinteo

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Posted 06 July 2002 - 12:18 PM

Qiagen kit .. is the best to extract Human DNA

#3 scbody

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Posted 06 July 2002 - 01:03 PM

Thank you

#4 FionaGould

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Posted 13 July 2002 - 04:35 AM

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.

#5 karen pooley

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Posted 13 July 2002 - 04:54 AM

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

kap28@mole.bio.cam.ac.uk


#6 scbody

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Posted 13 July 2002 - 01:12 PM

Karen and Fiona
Thank you for your input.
Regards
Simon






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