urgent help please

chiranjit chowdhury cjtcdy at gmail.com
Sun May 28 12:25:48 EST 2006


Hi!!

I am a research scholar of dept of Biotechnology ,IIT
Kharagpur,India.Myexperiment simply involves growing E coli containing
a pPBP fusion
construct,lysing the cell,purify the protein.Now I don't have access to a
sonicator.So I simply followed the protocol recommended by New England
Biolabs and prepared a lysis buffer containing lysozyme,EDTA,Tris and NaCl
(pH8).The volume of this buffer is 50 ml.But the main problem I have faced
is the suspension becomes highly viscous due to chromosomal DNA and it
becomes very hard to be taken out.I know that DNase could solve the problem
but I don't know how much should I have to add.  My DNase I (RNase free)conc
is 1U/ul.

With worm regards..



chiranjit chowdhury

research scholar

iit kharagpur

India


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