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Title
Culturing the colonies selected with Amp from the transformation done on July the 27th. We had 4 Amp plates:
- PMP: Visible colonies
- PGL 4.30: Visible colonies
- PHY32 (melanopsin): Nothing!
- - Control: Nothing
Since there was nothing in the PHY32 plates, we cheched the DNA concentration with Nanodrop (average over 2 samples, both very similar):
- 33.4 ng/µl of DNA
- 260/230 : 0.33
- 260/280 : 0.87
There is lots of stuff and not much DNA!!
We went ahead with the other 2 plasmids:
- Added 40 µl of Amp (100 µg/µl) to 40 ml of LB: 4000 µg / 40 ml = 100 µg/ml
- Distributed in 6 tubes, 5 ml each.
- Picked 3 colonies from each plate, PMP and PGL.
- Left overnight (starting at 19:30) at 37ºC.
Protocol: Prepare Plasmid Extraction (culture for Miniprep)
- Select and number colonies on the plates.
- Prepare tubes of LB medium with the correct quantity of antibiotics (100 µg/ml for Amp, Spc or chloramphenicol).
- Amp can be found in the -20ºC freezer at Ecoli, labeled as "stock". It is 100 µg/µl, or 1000x.
- The tubes to be used are the 14 ml round bottom found in front of the iGEM drawers (Falcon). Culture with cap in the first step (loose) and close to the second step after culture.
- Touch each colony with a clean pipette tip and put it in a tube.
- Put in incubator.
- Comments
Insert comments about what happened.
