using cheek cell dna

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solong
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using cheek cell dna

Post by solong » Tue Jul 31, 2007 6:36 pm

Anyone with workable molecular lab protocols using dna from cheek cells.
I work with it last semester but results were not so favorable.
Any suggestion is appreciated.
solong

sdekivit
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Post by sdekivit » Wed Aug 01, 2007 7:46 am

i did it a year ago. here's the protocol

1. vigorously scrape the inside of the cheek with a toothpick

2. place the toothpick in a clean microcentrifuge tube and add 1 mL water. Vortex 10 seconds and spin in a centrifuge 45 seconds at 10.000 rpm.

3. remove the toothpick and do not disturb the pellet! if pellet is distubed: centrifuge again.

4. remove 770 uL water. do not disturb the pellet.

5. add a little amount of chelex with a yellow tip.

6. vortex for 10 sec.

7. add 2 uL proteinase K and agitate.

8. incubate at 56 C for 5 minutes.

9. vortex for 10 seconds.

10. incubate in boiling water bath for 8 minutes.

11. vortex 10 seconds.

12. spin in a microcentrifuge at maximum speed for 3 minutes.

13. bottom = chelex and extraneous cellular matter, top layer = DNA. Sample = ready for PCR or may be stored at 4 C or frozen at -20 C. When reused after storage, thaw and repeat steps 12 and 13.

solong
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Post by solong » Wed Aug 01, 2007 7:27 pm

Thanx. I'll try this

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