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Miniprep for pMP and PGL4.30
- Cultures transfered to Falcon tubes and cetrifuged:
- pMP: 3 samples (1 to 3)
- pGL: 3 samples (1 to 3)
Protocol followed:
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
In the last centrifuge step, the cap of the pGL#2 tube got pulled off.
Then the 6 samples were checked with Nanodrop:
| sample | 260/280 | 260/230 | ng/µl |
|---|---|---|---|
| pMP#1 | 1.87 | 3.11 | 175.3 |
| pMP#2 | 1.87 | 3.10 | 162 |
| pMP#3 | 1.87 | 3.14 | 170.5 |
| pGL#1 | 1.86 | 2.72 | 229.5 |
| pGL#2 | 1.86 | 2.75 | 275 |
| pGL#1 | 1.86 | 2.69 | 251 |
