dna extraction

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Sat Nov 17 11:44:49 EST 2001


In <20011116192201.20298.qmail at ww02.jatek.com>, archna singh wrote:
>
>i have been trying to extract DNA from tissues/cells using the
>phenol/chloroform/isoamyl alcohol protocol.ive been using proteinase K at a
>temperature of 65 degrees for one hour prior to PCI extraction but the
>specific activity of the enzyme mentioned in the catalogue(Roche)is for 37
>degrees. im getting a 260/280 spectro reading of 1.2. is the proteinase K
>step likely to influence results so much?
>

No. The extinction coefficients for nucleotides are greater than those of most
proteins by at least one or two orders of magnitude (even at 280 nm).

More likely IMO is phenol contamination.

Nick

-- 
Nick Theodorakis
nicholas_theodorakis at urmc.rochester.edu

 




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