Hi,
I know there are some articles in this mail list about this issue, but ...
I am trying to perform homologous recombination on B. stearothermophilus. My
major problem is that I cannot transform the bugs! I tried the protoplast
protocol (Dr. Welker) with the NUB3621 strain, but did not have any success.
Even when using pUB110 as a positive control and different lysozyme
concentrations. I've been keeping the cells at high temperatures and
centrifuging at room temperature.
Does anybody out there have any ideas/advise on how to make this protocol to
work?
I am also about to give the electroporation protocol a try. Are there
anybody that have been working on that and can give some advise?
I very much appreciate any help.
Thanks,
Rafael
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