help needed with cosmid prep!!!

Mr. A.F. Hill ahill at crc.ac.uk
Wed Apr 7 09:59:04 EST 1993


hi

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,

thanks

Andy

==============================================================================
 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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