CTAB HELP

Helgi Briem Magnusson hbriem at rhi.hi.is
Wed Feb 9 08:46:55 EST 1994


In <ydegefu.12.2D579347 at viikki21.helsinki.fi> ydegefu at viikki21.helsinki.fi (YESHITILA DEGEFU (MMKAB_OPISK)) writes:

>Can anyone indicate a protocol for the use of CTAB in DNA purification? I 
>will be grateful if you also add your experience in using this reagent.

>Thank you. 

I have very good results with CTAB for purifying PCR template. 
Use CTAB stock of 10% CTAB-0.7M NaCl.  Heat in microwave.  This
solution does not flow at room temp, it is sort of jellylike.
When fluid, add 15ul to 100ul of NA prep.  This is rather difficult
to do accurately because of the viscosity.  Use a cut-off pipette
tip.  Mix well and let it stand on the bench for 20 min (I don't
know why). Spin in microfuge for 15-30 min depending on patience.
Discard the supernatant and resuspend the pellet in 50ul 1M NaCl
(I don't know why).  Add 500ul of EtOH.  Mix.  Microfuge 15 min
to pellet NA.  Resuspend.  CTAB is supposed to remove sugars.  It
has done wonders for me, but your mileage may vary.

Regards, Helgi Briem
Institute for Experimental Pathology
Keldur, Iceland



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