Melbourne/Lab Notebook

From 2007.igem.org

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(Week 2)
(Week 2)
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====Digest====
====Digest====
-
Digested
+
*[[Melbourne/Diagnostic Digest|Digested]] 5uL of each of the above miniprep DNA with EcoRI and PstI in buffer 3.
 +
*Incubated for 2hours 25min at 37degrees
 +
*Added 5uL 6x loading dye and stored at -20
 +
 
 +
====Glycerol Stocks====
 +
The following [[Melbourne/Glycerol Stocks|glycerol stocks]] were made:
 +
*Put aside from cultures 5/7 (labelled with this date)
 +
*#[[Melbourne/BBa_E0241|'''P2 15L 1''']]
 +
*#[[Melbourne/BBa_E0241|'''P2 15L 2''']]
 +
*#[[Melbourne/BBa_J61035|'''P4 8J 1''']]
 +
*#[[Melbourne/BBa_J61035|'''P4 8J 2''']]
 +
*#[[Melbourne/BBa_B0014|'''P1 1G 1''']]
 +
*#[[Melbourne/BBa_B0014|'''P1 1G 2''']]
 +
*Put aside from cultures 6/7 (labelled with this date)
 +
*#[[Melbourne/BBa_E0241|'''P2 15L 1''']]
 +
*#[[Melbourne/BBa_E0241|'''P2 15L 2''']]
 +
*#[[Melbourne/BBa_J61035|'''P4 8J 1''']]
 +
*#[[Melbourne/BBa_J61035|'''P4 8J 2''']]
 +
*[[Melbourne/BBa_E0040|'''P1 5H 1''']]
 +
*[[Melbourne/BBa_E0040|'''P1 5H 2''']]
 +
 
 +
====Liquid Culture====
 +
[[Melbourne/Growing up cells|Cultured]] the following in 5mL LB
 +
*[[Melbourne/BBa_I15010|'''I15010 1''']] (Kan)
 +
*[[Melbourne/BBa_I15010|'''I15010 2''']]
 +
*[[Melbourne/BBa_R0084|'''P1 11H 1''']] (Amp)
 +
*[[Melbourne/BBa_R0084|'''P1 11H 2''']]
 +
 
 +
Also placed Kan plate in the incubator to test antibiotic efficiency - no growth on 8/7
 +
 
 +
===7 July 2007===
 +
 
 +
===8 July 2007===
==Week 3==
==Week 3==

Revision as of 23:11, 8 July 2007

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Contents

Week 1


  • 26 June 2007: Streaked the following cells:
    1. pJS010 (from solid agar, Amp)
    2. Fusion protein (from glycerol stock, Amp?)
    3. BBa_I15010 (from solid agar, Kan)


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

  • 3 July 2007: Re Transformed into Joe's competent DH5alpha cells. Each Plate had additional 40uL of 50mg/ml Ampicillan spread on surface.
    1. P4 8J -> Three colonies -> grew in liquid culture 4 july
    2. P1 5H -> Multiple colonies at edge -> did not grow in overnight liquid culture 4 july 'poor amp spreading'
    3. P2 15L -> Three colonies -> grew in liquid culture 4 july
  • 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

Digest

Performed the following digests on DNA from the above miniprep

EcoRI/PstI with buffer 3
EcoRI/HaeII in buffer 2
XbaI/SpeI in buffer 2

Incubated for 2hours at 37 degrees, before addition of loading dye and storage at -20

Transformation

Liquid Culture

6 July 2007

Digest Gel

Miniprep

  • Put aside 1mL of liquid cultures set up on the 5th for glycerol stocks and labelled with todays date 6/7
  • Miniprepped the remains of the cultures and labelled with todays date 6/7. Samples were eluted with TE buffer.
  • P1 5H 1
  • P1 5H 2
  • P4 8J 1
  • P4 8J 2
  • P2 15L 1
  • P2 15L 2


Digest

  • Digested 5uL of each of the above miniprep DNA with EcoRI and PstI in buffer 3.
  • Incubated for 2hours 25min at 37degrees
  • Added 5uL 6x loading dye and stored at -20

Glycerol Stocks

The following glycerol stocks were made:

Liquid Culture

Cultured the following in 5mL LB

Also placed Kan plate in the incubator to test antibiotic efficiency - no growth on 8/7

7 July 2007

8 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