Performed a miniprep on the R0010 + E0241 inoculations from 6/19.
Created glycerol stocks of two cultures.
Creating Glycerol Stocks Protocol
- Obtain a screwcap-top eppendorf tube.
- Add 100 µL 50% glycerol.
- Add 100 µL liquid culture.
When the 2 colonies were centrifuged the pellets fluoresced green.
Made minipreps of the two pelleted cultures using the Qiagen protocol.
Performed digest on the R0010 + E0241 using XbaI and PstI
- Create 4 µL stock of 1 µL Xba + 1 µL Pst1 + 2 µL dH20 (Restriction Enzyme Stock)
- Create 4 µL stock of 1 µL BSA + 3 µL dH20 (BSA Stock)
- Pipet the following into the two tubes:
- 8 µL DNA
- 2 µL Restriction Enzyme Stock
- 1 µL BSA Stock
- 2.5 µL buffer 3
- 11.5 µL dH20
Total volume: 25 µL
- Incubate at 37 Celcius for 1 hour
Jeff took care of the digestions after incubation: I left the lab at 12:30 PM.I went on a trip to Torrance, CA, to visit my sister's graduation.