Biolistic Nuclear transformation

Andy Vaughan fleaghost at hotmail.com
Fri Aug 16 10:53:03 EST 2002


Hello all,

I am attempting some experiments in which I need to do nuclear 
transformation with the PDS-1000/He gene gun of chlamy.  I have done a large 
amount of shooting, but have had no good results.  I've tried shooting on 
cells imbedded in agar and agarose, cells actually grown up on the plates, 
and cells on nylon membranes (the best candidate thus far), but have had no 
success.  I've also tried a variety of pressure disks and target locations, 
macrocarrier locations, and microparticle preparation methods.  I have tried 
both the pJD67 (arg plasmid) and pSP124s (ble plasmid), but have had no 
discernable transformants with either.  I'm starting to get pretty 
desperate, so any help would be greatly appreciated.  Thanks.


Sincerely,

Andy Vaughan
fleaghost at hotmail.com




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