Melbourne/Lab Notebook

From 2007.igem.org

(Difference between revisions)
(27 June 2007)
(27 June 2007)
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**Used resuspended DNA that was stored on Monday
**Used resuspended DNA that was stored on Monday
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=====Liquid culture=====
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====Liquid culture====
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#[[Melb:Growing Cells|Prepared]] 30mL of Amp LB (100ug/mL). Prepared 5mL Kan LB (50mg/mL).
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#[[Melb:Growing up cells|Prepared]] 30mL of Amp LB (100ug/mL). Prepared 5mL Kan LB (50mg/mL).
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#Aliquoted 5mL Amp LB into 6 50mL falcon tubes
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#To the Amp LB the following were introduced:
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*
===28 June 2007===
===28 June 2007===

Revision as of 01:39, 3 July 2007

<Back to team home page>

Contents

Week 1

25 June 2007

Prepared LB agar plates.

Transformation

  1. resuspended the following from Registry plates:
    • BBa_I15008 - ho1 (P2 21A, Kan)
    • BBa_I15009 - PcyA (P2 21C, Kan)
    • BBa_R0084 - OmpR positive promoter (P1 11H, Amp)
    1. Punctured foil with pipette tip.
    2. Resuspended in 15uL ddH2O.
    3. Stored in-20 (after taking 1uL for transformation).
  1. Transformed into competent DH5alpha cells with shorter incubation times as follows:
    • 30min on ice after DNA addition
    • 10min on ice after heat shock
    • 30min at 37degrees with LB

26 June 2007

Transformation from Monday

  • Transformation of BBa_I15008 and BBa_I15009 failed. No colonies on plates
  • Small number of colonies on BBa_R0084 plate.
  • Placed in cool room

Transformation

  1. Resuspended the following parts in 15uL:
  • BBa_B0034 (RBS, plate 1 3O, Amp)
  • BBa_C0051 (C1 repressor, plate 1 5G, Amp)
  • BBa_B0010 (Terminator, plate 2 3P, Amp)
    • Think some DNA may have remained in wells
  1. Transformed into competent DH5alpha cells from Joe

Streak plates

  1. Streaked the following cells:
  • PJS010 (from solid agar, Amp)
  • Fusion protein (from glycerol stock, Kan)
  • BBa_I15010 (from solid agar, Kan)

27 June 2007

Transformation

  1. Repeated transformation of failed parts from Monday:
  • BBa_I15008 (Kan)
  • BBa_I15009 (Kan)
    • Used resuspended DNA that was stored on Monday

Liquid culture

  1. Prepared 30mL of Amp LB (100ug/mL). Prepared 5mL Kan LB (50mg/mL).
  2. Aliquoted 5mL Amp LB into 6 50mL falcon tubes
  3. To the Amp LB the following were introduced:

28 June 2007

29 June 2007

30 June 2007

Week 2

2 July 2007

Now

3 July 2007

4 July 2007

5 July 2007

6 July 2007

Week 3

9 July 2007

10 July 2007

11 July 2007

12 July 2007

13 July 2007

Week 4

16 July 2007

17 July 2007

18 July 2007

19 July 2007

20 July 2007

Week 5

23 July 2007

24 July 2007

25 July 2007

26 July 2007

27 July 2007

Week 6

30 July 2007

31 July 2007

1 Aug 2007

2 Aug 2007

3 Aug 2007