Preparation of pUC 19 (Done with Hank)
- Took out ON culture of pUC19-containing cells (innoculated on 12 June 2009).
- Note that the culture rotor was not spinning - tube may not have been well-aerated.
- Mixed the entire 80µL of RNaseA into Charge-Switch Pro Resuspension buffer.
- Aliquoted 1.25 mL of ON culture each per microfuge tube (4 in total), spun down at max speed for 10 min.
- Followed ChargeSwitch Pro miniprep protocol from here on; combined the harvested cells into two tubes (one from Hank, one from me).
- Nanodropped the samples:
| pUC19 (1)
| pUC19 (2)
- Stored at AMBLE -20ºC freezer
- Moved to Lagally Lab -20ºC freezer, labelled:
- pUC19 HYEM(1)
- pUC19 HYEM(2)
- Made glycerol stocks of these cells (250µL of 60% sterile glycerol + 750µL of ON culture)
Results of Mutagenesis
- Hank noticed that the media was cracked; could be too thin.
- Took plates out from AMBL 37ºC incubator @ 2:45 PM.
| Positive (conc.)
| Positive (dil.)
| Negative (conc.)
| Negative (dil.)
| Control (dil.)
| Control (conc.)
- Plates were parafilmed and stored at AMBL4ºC.