l**********8
发帖数: 2 | 1 Hi, are there anyone familiar with E coli?
I've screened some mutants and is trying to confirm them by doing site
mutagenesis and spot titration. But, during the process I found my
transformants which were streaked out twice, were contaminated by phage (
precipitates and clear o/n culture without many cells). When I am doing the
o/n culture, I found that if I add Kan, there would be a phage problem, but
if I didn't, there wouldn't. This makes me sooooooo confused and
disappointed. I wonder whether there are anybody who work with bacteria can
give me some suggestions. Thank you!
P.S.
I am now growing my cells in 37C with or without Kan at the same time to
figure out wheter it is antibiotics inducible. | b******y
发帖数: 627 | 2 My guess,
Kan slow down the growth a bit. That allows phage to catch up and clear out
your culture.
Can you use TonA knockout strain to do your experiments? if so, you can try
that. (You can get the strain from a number of vendors.) The reason is that
the popular bacteria phage contamination is due to T1 phage. The receptor of
it on E. coli is TonA.
Good luck. | l**********8
发帖数: 2 | 3 Thank you so much!
My today's experiment also shows that after adding Kan, my bacteria got a
phage problem probably due to the reason you mentioned above.
out
try
that
of
【在 b******y 的大作中提到】
: My guess,
: Kan slow down the growth a bit. That allows phage to catch up and clear out
: your culture.
: Can you use TonA knockout strain to do your experiments? if so, you can try
: that. (You can get the strain from a number of vendors.) The reason is that
: the popular bacteria phage contamination is due to T1 phage. The receptor of
: it on E. coli is TonA.
: Good luck.
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