Help: genomic DNA refused to be cut

Li Bing sar96lb at reading.ac.uk
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                               
UK
Tel. 01734-875123 ext.7025 (lab.)
--------------------------------------




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