cry transformation change

[Julie Nelson]

I have discovered a modification to my transformation protocol using 
pJN4, which is the CRY1-1 gene with a truncated RBCS2 promoter, which 
both improves efficiency and saves time (see Mol. Cell. Biol. 14: 
4011-4019, transformation method 3, for original protocol).  Leave out 
the step of eight hours in SGII right before plating.  So after 
transformation, the steps should be:  3.5 hours in 10 ml SGII, 4 days 
in 100 ml SGII-N, pellet cells, spread onto emetine plates.  The 
efficiency improved about two-fold for me.

Julie Nelson
julie at molbio.cbs.umn.edu