Melbourne/Lab Notebook
From 2007.igem.org
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====Miniprep==== | ====Miniprep==== | ||
| - | *miniprepped the following overnight cultures set up on the 4th of July | + | *miniprepped the following overnight cultures set up on the 4th of July. Final elution was performed with TE buffer prepared on 25/06/07 rather than nuclease free water. |
**[[Melbourne/BBa_B0010|'''P2 3P 1''']] | **[[Melbourne/BBa_B0010|'''P2 3P 1''']] | ||
**[[Melbourne/BBa_B0010|'''P2 3P 2''']] | **[[Melbourne/BBa_B0010|'''P2 3P 2''']] | ||
| Line 148: | Line 148: | ||
**[[Melbourne/BBa_E0241|'''P2 15L 1''']] | **[[Melbourne/BBa_E0241|'''P2 15L 1''']] | ||
**[[Melbourne/BBa_E0241|'''P2 15L 2''']] | **[[Melbourne/BBa_E0241|'''P2 15L 2''']] | ||
| - | **[[Melbourne/BBa_Q04510|'''P2 13K 1''']] | + | **[[Melbourne/BBa_Q04510|'''P2 13K 1''']](eluted in 130uL due to accidental double application of 50uL elution) |
**[[Melbourne/BBa_Q04510|'''P2 13K 2''']] | **[[Melbourne/BBa_Q04510|'''P2 13K 2''']] | ||
**[[Melbourne/BBa_B0014|'''P1 1G 1''']] | **[[Melbourne/BBa_B0014|'''P1 1G 1''']] | ||
Revision as of 02:24, 5 July 2007
Contents |
Week 1
25 June 2007
Prepared LB agar plates.
Transformation
- resuspended the following from Registry plates:
- Punctured foil with pipette tip.
- Resuspended in 15uL ddH2O.
- Stored in-20 (after taking 1uL for transformation).
- 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
- Resuspended the following parts in 15uL:
- Think some DNA may have remained in wells
- Transformed into competent DH5alpha cells from Joe
Streak plates
- Streaked the following cells:
- pJS010 (from solid agar, Amp)
- Fusion protein (from glycerol stock, Amp?)
- BBa_I15010 (from solid agar, Kan)
27 June 2007
Transformation
- Used resuspended DNA that was stored on Monday
Liquid culture
- Prepared 30mL of Amp LB (100ug/mL). Prepared 5mL Kan LB (50mg/mL).
- Aliquoted 5mL Amp LB into 6 50mL falcon tubes
- To the Amp LB aliquots single transformed colonies from plates of the following were introduced:
- To the Kan LB a single colony from the transformation plate of BBa_I15010 was introduced.
- Cells incubated at 37degrees with shaking overnight.
28 June 2007
Miniprep
- Minipreped the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
- Stored in -20 freezer
Digest
Liquid culture
- Cultured the following cells from transformed plates:
29 June 2007
Miniprep
- Minipreped the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
- 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.
Liquid Culture
- Cultured 2 colonies from each of the following transformed plates and labelled as follows
5 July 2007
Miniprep
- miniprepped the following overnight cultures set up on the 4th of July. Final elution was performed with TE buffer prepared on 25/06/07 rather than nuclease free water.
- the following liquid cultures were not miniprepped due to failure (no growth)
