Virginia Commonwealth/22 July 2009

From 2009.igem.org

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** 1 mL of sample was diluted with 6 mL of LB + Cm
** 1 mL of sample was diluted with 6 mL of LB + Cm
* Samples from cryogenic stocks were grown up in LB + Cm:
* Samples from cryogenic stocks were grown up in LB + Cm:
 +
{| cellpadding="10" align="center" border="0.9" padding="5"
 +
|+
 +
|-
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! Promoter !! RET !! Growth Observation !!
 +
|-
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! J23100
 +
| seed stock|| LB || make more seed stock ||
 +
|-
 +
! J23101
 +
| electro comp. || LB || (+) Control/ make more stock ||
 +
|-
 +
! J23101 
 +
| electro comp. || LB || (+) Control shocked ||
 +
|-
 +
! J23102
 +
| electro comp. || LB || (+) Control shocked ||
 +
|-
 +
! J23102
 +
| electro comp. || LB || (+) Control/make more stock ||
 +
|-
 +
! J23103
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23103
 +
| electro comp. || Cm || J23100 w/ J06702 ||
 +
|-
 +
! J23104
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23104
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23105
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23105
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23106
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23106
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23107
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23107
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23110
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23110
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23113
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23113
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23116
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! J23116
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! R0011
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! R0011
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! R0040
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! R0040
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
 +
|}
 +
** J23100 w/ E0240 (taken from plates)
** J23100 w/ E0240 (taken from plates)
** J23101 w/ E0240
** J23101 w/ E0240

Revision as of 20:35, 22 July 2009

Contents

Wednesday 22 July 2009

Results

  • Overnight cultures were successful
    • 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

Trentay

Tasks

  • 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)

Wetlab

  • Dilute overnight cultures
    • 1 mL of sample was diluted with 6 mL of LB + Cm
  • Samples from cryogenic stocks were grown up in LB + Cm:
Promoter RET Growth Observation
J23100 seed stock LB make more seed stock
J23101 electro comp. LB (+) Control/ make more stock
J23101 electro comp. LB (+) Control shocked
J23102 electro comp. LB (+) Control shocked
J23102 electro comp. LB (+) Control/make more stock
J23103 electro comp. Cm (-) Control
J23103 electro comp. Cm J23100 w/ J06702
J23104 electro comp. Cm (-) Control
J23104 electro comp. Cm (-) Control
J23105 electro comp. Cm (-) Control
J23105 electro comp. Cm (-) Control
J23106 electro comp. Cm (-) Control
J23106 electro comp. Cm (-) Control
J23107 electro comp. Cm (-) Control
J23107 electro comp. Cm (-) Control
J23110 electro comp. Cm (-) Control
J23110 electro comp. Cm (-) Control
J23113 electro comp. Cm (-) Control
J23113 electro comp. Cm (-) Control
J23116 electro comp. Cm (-) Control
J23116 electro comp. Cm (-) Control
R0011 electro comp. Cm (-) Control
R0011 electro comp. Cm (-) Control
R0040 electro comp. Cm (-) Control
R0040 electro comp. Cm (-) Control
    • J23100 w/ E0240 (taken from plates)
    • J23101 w/ E0240
    • J23101 w/ J06702
    • J23102 w/ E0240
    • J23102 w/ J06702
    • J23103 w/ E0240
    • J23103 w/ J06702
    • J23104 w/ E0240
    • J23104 w/ J06702
    • J23105 w/ E0240
    • J23105 w/ J06702
    • J23106 w/ E0240
    • J23106 w/ J06702
    • J23107 w/ E0240
    • J23107 w/ J06702
    • J23110 w/ E0240
    • J23110 w/ J06702
    • J23113 w/ E0240
    • J23113 w/ J06702
    • J23116 w/ E0240
    • J23116 w/ J06702
    • R0011 w/ E0240
    • R0011 w/ J06702
    • R0040 w/ E0240
    • R0040 w/ J06702
  • 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
  • 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 20:27, 22 July 2009 (UTC)

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