Virginia Commonwealth/22 July 2009

Wednesday 22 July 2009

Maria and Afton

Overnight cultures were successful
- However, vials had noticeable amounts of white E. coli at the bottom of the tube.
Flow Cytometer was repaired
- Samples cannot be tested until protocols for using and maintaining the machine are figured out
Plated bacteria are still not expressing GFP and RFP

Maria and Afton

Dilute E. coli into fresh LB + Cm in preparation for testing with the flow cytometer
- nothing was tested today with the cytometer
Grow all BioBrick parts in 5 mL of LB + Cm overnight
- 100 microliters was added to broth
- plastic tubes had tops, so tops were loosened during the incubation period
- cells were out of the -80 degree Celsius freezer for a prolonged amount of time, so viability is lower than usual.
  - If nothing grows, BioBrick DNA will be reinserted into fresh NEB 10 beta through electrotransformation.
Retransform J23100 w/ J06702
Transform NEB 10 beta and DB 3.1 to check for electrocompetency
Organize notebook and running draft

Trentay 19:57, 22 July 2009 (UTC)

Maria and Afton

Dilute overnight cultures
- 1 mL of sample was diluted with 6 mL of LB + Cm
- only seven samples were saved however, most of these parts were recultured and the flow cytometer was not used today, so these cell cultures most likely will not be kept
Samples from cryogenic stocks were grown up in LB:
- DB 3.1 (2 vials)
Samples from cryogenic stocks were grown up in LB + Amp:
- pSB4C5
Samples from cryogenic stocks were grown up in LB + Cm:

Transformation of electrocompetent E. Coli stock and BioBrick with promoter J23100 and RET J06702


Name	Type	Plate	Purpose
NEB 10 beta	seed stock	LB	make more seed stock
NEB 10 beta	electro comp.	LB	(+) Control/ make more stock
NEB 10 beta	electro comp.	LB	(+) Control shocked
DB 3.1	electro comp.	LB	(+) Control shocked
DB 3.1	electro comp.	LB	(+) Control/make more stock
NEB 10 beta	electro comp.	Cm	(-) Control
NEB 10 beta	electro comp.	Cm	J23100 w/ J06702

Trentay 20:51, 22 July 2009 (UTC)