Login

|
|
using cheek cell dnaModerator: BioTeam
3 posts • Page 1 of 1
using cheek cell dnaAnyone 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
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.
3 posts • Page 1 of 1
Who is onlineUsers browsing this forum: No registered users and 0 guests |
© Biology-Online.org. All Rights Reserved. Register | Login | About Us | Contact Us | Link to Us | Disclaimer & Privacy
Science Network - Braintrack.com - University Directory | Chemicool.com - Chemistry | Logo design by LogoBee | Powered by phpBB