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PCR Cleanup

We use primarily ExoSAP-IT© for PCR Product Clean-Up PDF for standard cleanup of successful PCR.

Alternatively we occasionally use a gel purification and isolation technique in which case we use the Gene Clean method
  1. Run standard gel (~1.5%) with 1X TAE (TBE does not work as well)
  2. Cut out band using clean razor blade
  3. Weigh agarose in mg
  4. Add 3X agarose weight in microliters of NaI
  5. Incubate 5-10 min at 55oC until agarose is thoroughly dissolved
  6. Suspend Glass Milk by thorough vortexing
  7. Add 5 l Glass Milk to agarose solution
  8. Mix by inverting and vortexing
  9. Place on ice for 5 min vortexing about every 2 min
  10. Centrifuge tube on high 5 sec
  11. Pipette off supernatant
  12. Keep tubes on ice as much as possible. Add 600 l of cold New Wash, and resuspend pellet by vortexing. (note: store New Wash tightly sealed at -20ºC)
  13. Centrifuge tube on high 15 sec
  14. Pipette off supernatant
  15. Go to step 12 two more times
  16. Air dry for 15-20 min
  17. Resusped in 12 l TE buffer or ultra-pure H2O
  18. Incubate ate 45 -55oC for 3 min
  19. Centrifuge 30 sec
  20. Remove supernatant (with DNA) into clean tube

 
 

Kelly Miller Lab, Department of Biology, University of New Mexico, 167 Castetter Hall, MSC03 2020, Albuquerque, NM 87131-0001 USA
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