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Miniprep
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 protocol (page 22) and a microcentrifuge.
- Comments
The bacteria were incubated in 15ml tubes with 5ml medium and transferred to two 2ml microcentrifuge tubes.
The column 2* was spilled before the first centrifugation step.
Two types of Ampicillin were used, the rests of last year's iGEM team and the lab's supply. The tubes 1 and 2 were made using the old one, tubes 3, 4 and 5 using the lab's Amp (this was done to check if the old Amp still worked).
Aliquoted all 5 of them & froze them at -20°C.
