I am trying to make DH1 chemical competent cells using the transformation buffer found here
. However, my cells look very flaky and clumpy when I try to resuspend them in the transformation buffer. I'm not sure if it's the buffer, the nature of DH1 or something that I've done wrongly. I streaked DH1 cells on LB-agar with 15ug/mL nalidixic acid, then inoculated a colony into 10mL LB with 15ug/mL nalidixic acid for overnight growth at 37C. 1 mL of the overnight cells was then added to 400mL of LB and grown to OD600 0.4. The cells were contrifuged at 4000g for 3min in 50mL falcon tubes. This was when I added the transformation buffer to the pellet and had difficulty resuspending the cells. I tried TSS buffer
and got the same results.
Has anyone have such experience too?
EDIT: Just to add that I just took the clumpy part and diluted with water. They seem to resuspend very well. What's going on?!?!
Edited by donny, 02 August 2012 - 06:21 PM.