Help: genomic DNA refused to be cut

Li Bing sar96lb at
Mon Mar 9 13:42:29 EST 1998

Hi all,

I`m looking for help to make my genomic DNA cut with restriction enzymes.

I`m working on crab genomic DNA which seems determined not be cut with any
enzymes. I asked help before in this newsgroup and I got some respond.

This is the list of protocols I tried:

*Extra phenol and chloroform extractions
*Different protocols using CTAB then phenol/chloroform
*Guanidine thiocyanate extraction(the same buffer as used for RNA but
using PH8 phenol instead)
*Dialysis against TE buffer for 72 hours, changing buffer 3 times

Many thanks for you friends who sent me all this protocols. 

Pity me, I still can`t cut my dear crab DNA although 5 crabs sacrificed.
I don`t want kill any crab for that purpose anymore. So if there is anyone
who got a reliable method, Please, Please let me know. Or there will be
other guys keep trying to make it work. This means more crabs will be
forced  diving into hot water...
Li Bing                               
School of Animal & Microbial Science  
The University of Reading             
Whiteknights, PO Box 228              
Reading, Berkshire                    
RG6 6AJ                               
Tel. 01734-875123 ext.7025 (lab.)

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