"
Morning
- Comments
Took the falcon tubes out of the incubator. There was growth for all concentrations of Spectinomycin (Spc) (slightly less for the higher concentrations, but the difference is not easily noticeable.
Our competent cells grew very well on the ampicillin plates, but nothing grew on the Spc plates.
Successfully prepared the competent cells for the mini-prep in the afternoon (bacteria have been pelleted and put in the freezer).
Afternoon
Protocol: Miniprep
The slim tubes can be centrifuged in the machine in front of the "Gel hood", at 4000 rpm for 10 min. The fatter ones, in the E. coli centrifuge by the fridge (the tip can be left inside, since it floats).
Pellets resuspended with RNase containing buffer (Resuspension Buffer R3, from Invitrogen, equivalent to Buffer P1 from Qiagen, in Sowmya's box in the fridge). Note: keep the buffer in ice if you are not bringing it back to the fridge for some minutes.
We then use the QIAGEN QIAprep Spin Miniprep Kit with their [http://www.qiagen.com/literature/render.aspx?id=370 protocol] (page 22) and a microcentrifuge.
- Comments
This was done with the LovTAP Read-Out plasmid. The QC hasn't been done yet.
