Melbourne/Lab Notebook
From 2007.igem.org
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==Week 1== | ==Week 1== | ||
| - | + | *25 June 2007: | |
| + | **Prepared LB agar plates Amp & Kana. | ||
| + | **[[Melbourne/IGEM2007 kit|resuspended the following from Registry plates]] & [[Melbourne/Transformation Protocol shorter|Transformed (shorter protocol)]] into competent DH5alpha cells. | ||
| + | **#[[Melbourne/BBa_I15008|'''P2 21A''']] - (BBa_I15008, ho1, Kan) | ||
| + | **#[[Melbourne/BBa_I15009|'''P2 21C''']] - (BBa_I15009, PcyA, Kan) | ||
| + | **#[[Melbourne/BBa_R0084|'''P1 11H''']] - (BBa_R0084, OmpR positive promoter, Amp) | ||
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| - | + | *26 June 2007 | |
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====Transformation from Monday==== | ====Transformation from Monday==== | ||
Revision as of 04:48, 5 July 2007
Contents |
Week 1
- 25 June 2007:
- Prepared LB agar plates Amp & Kana.
- resuspended the following from Registry plates & Transformed (shorter protocol) into competent DH5alpha cells.
- 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.
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
- 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)
