Help: genomic DNA refused to be cut

[Mathieu Ingouff]

Dear Li,

I have just tried a protocol (NAR, 1997, vol 25, no 25) that seems to be
universal as they said : Wheat, Barley, Shrimps, Eucalyptus...
you may want to kill another crab to try it?
In my case it works for spruce.


Mathieu



>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.)
>--------------------------------------




____________________________________
  Mathieu Ingouff, PhD student
  SLU, Department of  Forest Genetics
  Genetikv=E4gen 1B, Box 7027
  S-750 07 Uppsala, Sweden

  email: mathieu.ingouff at sgen.slu.se
  fax: +46.18.67.32.79.
 phone: +46.18.67.32.32.
____________________________________