Virginia Commonwealth/22 July 2009

From 2009.igem.org

(Difference between revisions)
(→Wednesday 22 July 2009)
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** pSB4C5
** pSB4C5
* Transformation of electrocompetent E. Coli stock and BioBrick with promoter J23100 and RET J06702
* Transformation of electrocompetent E. Coli stock and BioBrick with promoter J23100 and RET J06702
-
** NEB 10 beta (seed stock)    plated on LB
+
{| cellpadding="10" align="center" border="0.9" padding="5"
-
** NEB 10 beta (electro comp.) plated on LB
+
|+
-
** NEB 10 beta (electro comp.) plated on LB (shocked)
+
|-
-
** DB 3.1     (electro comp.) plated on LB (shocked)
+
! Name !! Type !! Plate !! Purpose !!
-
** DB 3.1     (electro comp.) plated on LB
+
|-
-
** NEB 10 beta (electro comp.) plated on Cm  
+
! NEB 10 beta  
-
** NEB 10 beta (electro comp.) J23100 w/ J06702 plated on Cm
+
| seed stock|| LB || make more seed stock ||
 +
|-
 +
! NEB 10 beta  
 +
| electro comp. || LB || (+) Control ||
 +
|-
 +
! NEB 10 beta
 +
| electro comp. || LB || (+) Control shocked ||
 +
|-
 +
! DB 3.1  
 +
| electro comp. || LB || (+) Control shocked ||
 +
|-
 +
! DB 3.1  
 +
| electro comp. || LB || (+) Control ||
 +
|-
 +
! NEB 10 beta  
 +
| electro comp. || Cm || (-) Control ||
 +
|-
 +
! NEB 10 beta  
 +
| electro comp. || Cm || J23100 w/ J06702 ||
 +
|}

Revision as of 20:25, 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:
    • 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
NEB 10 beta electro comp. LB (+) Control shocked
DB 3.1 electro comp. LB (+) Control shocked
DB 3.1 electro comp. LB (+) Control
NEB 10 beta electro comp. Cm (-) Control
NEB 10 beta electro comp. Cm J23100 w/ J06702
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