Kiwi DNA

arnaud.amelot at libertysurf.fr arnaud.amelot at libertysurf.fr
Sat Mar 3 06:24:24 EST 2001


Hello, my name is Aymeric, I study Biochemistry at the
university of LILLE I in France.
I’m now working on Kiwi DNA and I have two problems in my
protocol to isolate this DNA:
   1°, I can gather only a very little quantity of DNA.
   2°, Moreover in final, this little quantity DNA that
obtain is not pure, I also get a lot of peptine that I
Would want evacuate
 This is my protocol:
 Tissue dilacerations.
 Cells are burst with 15 ml of plug: Tris/HCL 50 mM, pH
8, NaCL100 mM, 20 mM EDTA, 1% SDS.
 After fidgetiness during 5 minutes, add 15 ml of
saturate phenol pH 8.
 Centrifugation during 5 minutes to 3000 rpm.
 The upper phase containing DNA is deducting in, adding
of 1 volume of Chloroform / isoamylic alcohol, waved
during 10 minutes, following of a new centrifugation ( 5
minutes to 3000 rpm).
 The upper phase is deducting in, and DNA is precipated
with 1 volume of ethanol.
  DNA precipated is rolling up and redissolved in 5 ml of
plug: Tris/ HCL 10 mM pH            7,4, NaCL 10 mM, EDTA
1 mM , after  to be dialysed in the same Plug.

Thanks a lot for your help, this is my e-mail adress:
aymamelot at hotmail.com


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