help needed with cosmid prep!!!
Mr. A.F. Hill
ahill at crc.ac.uk
Wed Apr 7 09:59:04 EST 1993
ive been trying to grow up a cosmid (~45kb) and have had varying results
using different methods. the problem is that if i get DNA i cant cut it with
restriction enzymes. i have used the standard alkaline lysis / PEG precipitn
methods and also the triton lysis method. i can cut the dna from peg pptn
but from a 500ml prep i only get 1-5 ug DNA which is a pretty low yield.
ive noted that most of the dna is lost at the peg/nacl pptn stage as when
i reprecipitate the supernatant there is heaps of dna left - any reason
why this is happening?
hope this makes sense to someone!
any suggestions welcomed,
Andy Hill |
Prion Disease Group | E-Mail: ahill at uk.ac.crc
Dept. Biochemistry & Molecular Genetics |-----------------------------------
St Mary's Hospital Medical School | Phone: 071 723 1252 ext. 5469
LONDON W2 1PG | Fax: 071 706 3272
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