Melbourne/Lab Notebook

From 2007.igem.org

(Difference between revisions)
(Miniprep)
(4 July 2007)
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*Made up 25 Ampicillin plates using 400mL LB agar prepared on the 25th of June.   
*Made up 25 Ampicillin plates using 400mL LB agar prepared on the 25th of June.   
**Used 100mg/ml Ampicillin stock from Gooley Lab - In bag in left freezer near our lab bench.
**Used 100mg/ml Ampicillin stock from Gooley Lab - In bag in left freezer near our lab bench.
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 +
====Tranformation====
 +
[[Melbourne/Transformation Protocol|Transformed]] the following and grew on new ampicillin plates
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*[[Melbourne/BBa_E0040|'''P1 5H''']]
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*[[Melbourne/BBa_J61035|'''P4 8J''']]
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*[[Melbourne/BBa_E0241|'''P2 15L''']]
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**Also Plated an untransformed control (subjected to transformation protocol in absense of DNA) and a blank plate control (no cells)
====Liquid Culture====
====Liquid Culture====

Revision as of 03:23, 5 July 2007

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Contents

Week 1

25 June 2007

Prepared LB agar plates.

Transformation

  1. resuspended the following from Registry plates:
    • P2 21A - (BBa_I15008, ho1, Kan)
    • P2 21C - (BBa_I15009, PcyA, Kan)
    • P1 11H - (BBa_R0084, OmpR positive promoter, Amp)
  2. Punctured foil with pipette tip.
  3. Resuspended in 15uL ddH2O.
  4. Stored in-20 (after taking 1uL for transformation).
  5. Transformed into competent DH5alpha cells with shorter incubation times as follows:
    1. 30min on ice after DNA addition
    2. 10min on ice after heat shock
    3. 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:
    • P1 3O (BBa_B0034, RBS, Amp)
    • P1 5G (BBa_C0051, c1 protein, Amp)
    • P2 3P (BBa_B0010, Terminator, Amp)
  • Think some DNA may have remained in wells
  1. Transformed into competent DH5alpha cells from Joe

Streak plates

  1. Streaked the following cells:

27 June 2007

Transformation

  1. Repeated transformation of failed parts from Monday:
    1. P2 21A (Kan)
    2. P2 21C (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 aliquots single transformed colonies from plates of the following were introduced:
  4. To the Kan LB a single colony from the transformation plate of BBa_I15010 was introduced.
  5. Cells incubated at 37degrees with shaking overnight.

28 June 2007

Miniprep

  1. Minipreped the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
  2. Stored in -20 freezer

Digest

Liquid culture

  1. Cultured the following cells from transformed plates:

29 June 2007

Miniprep

  1. Minipreped the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
  1. Stored in -20 freezer

30 June 2007

Week 2

2 July 2007

Now

3 July 2007

4 July 2007

Ampicillin Plates

  • Made up 25 Ampicillin plates using 400mL LB agar prepared on the 25th of June.
    • Used 100mg/ml Ampicillin stock from Gooley Lab - In bag in left freezer near our lab bench.

Tranformation

Transformed the following and grew on new ampicillin plates

  • P1 5H
  • P4 8J
  • P2 15L
    • Also Plated an untransformed control (subjected to transformation protocol in absense of DNA) and a blank plate control (no cells)

Liquid Culture

5 July 2007

Miniprep

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