strange bacteria culture - (Oct/09/2008 )
Hi!
Could anyone make a quick guess on what went wrong.
I did normal cloning and got around 50 colonies on the plate and zero from the control (empty vector). These were Amp plates with about 50ug/ml antibiotic concentration.
Yesterday I set up miniprep and today morning I found that in all of the tubes (5ml Falcon tubes with LB-Amp at 50ug/ml) the bacteria are mostly at the bottom and the LB is more or less clear. Someone else is doing miniprep and gets a cloudy culture in the same shaker and same kind of tubes.
If I shake my tubes the LB gets very cloudy of course, but do you think the bacteria died? Why did they stick to the bottom? I am now doing the miniprep to find out, whether some have the right insert.
Cheers!
R
Could anyone make a quick guess on what went wrong.
I did normal cloning and got around 50 colonies on the plate and zero from the control (empty vector). These were Amp plates with about 50ug/ml antibiotic concentration.
Yesterday I set up miniprep and today morning I found that in all of the tubes (5ml Falcon tubes with LB-Amp at 50ug/ml) the bacteria are mostly at the bottom and the LB is more or less clear. Someone else is doing miniprep and gets a cloudy culture in the same shaker and same kind of tubes.
If I shake my tubes the LB gets very cloudy of course, but do you think the bacteria died? Why did they stick to the bottom? I am now doing the miniprep to find out, whether some have the right insert.
Cheers!
R
Don't worry. Perhaps u didn't have them shaking (hard enoguh) when culturing. How long was your culture, if it's more than like one week then might have some death occurring. if it's overnight .. then no.
Just go ahead and do your miniprep. relax.
Yes, I think maybe the shaking speed is not enough.
Thanks!
Whatever the reason was the miniprep came out OK and I got the correct insertion. 