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10-23-2003, 09:47 AM
  #5
Le Gross Pork
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vCash:
Firstly, we must take 3 things into consideration:

(I) Difficulty or success in floral transformation with Arabidopsis

(II) Problems of selecting for the pSOUP vector whose trfA function provides the replicase function for pGreen (see below). Many C58-derived strains (including GV series) develop spontaneous resistance to tetracyclin. The solution is to co-transform both pSoup and pGreen and use kanamycin selection solely, rather than tet/kan.
A chemical transformation method is available in MolBiol.Net > Protocols > Transformations

(III) Backbone rearrangements, pGreenI was prone to recombination with E.coli genomic DNA - has anyone has problems with the same and pGreenII