(no subject)

Sissi zhang via methods%40net.bio.net (by sissi_2009 from hotmail.com)
Mon Mar 2 20:44:11 EST 2009









Kyle

Thank you for the advice. I am still trying to figure out why my culture is not growing well. 
Now
I just test the starter culture since it did not grow to the expected
cell densities in the first place. The OD600=0.75 after 20 hr
incubation at 37C at 300 rpm. The culture was half transparent after 20
hrs growth.

Here was the procedure:
The colony was picked from fresh agar plate.
The single colony (around 1.2 mm in diameter) was inoculated to 2.5 mL LB broth (with or without Amp)
After 20 hrs growth, both tubes gave me the same cell density, OD600=0.75 (with or without 100 ug/mL Amp).
I checked the pH of LB broth and adjust it to pH7.5 before autoclave.

I
really do not know what the trouble is. Transformed DH5 +pUC19 should
grow very fast. I just placed an order for liquid LB broth from
different vendor (not dehydrated LB), in case the problem lies in the
water I used to make LB broth or something. Could it be contamination
issue? But I don't see  how. 

The super broth formula you used seems very good. I will try it if the new LB broth I just ordered will not work. Thanks, Sissi

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