Melbourne/Lab Notebook
From 2007.igem.org
(Difference between revisions)
| Line 3: | Line 3: | ||
==Week 1== | ==Week 1== | ||
*25 June 2007: Prepared LB agar plates Amp & Kana. | *25 June 2007: Prepared LB agar plates Amp & Kana. | ||
| - | *25 June 2007: [[Melbourne/IGEM 2007 kit| | + | *25 June 2007: [[Melbourne/IGEM 2007 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) -> No colonies on plates | *#[[Melbourne/BBa_I15008|'''P2 21A''']] - (BBa_I15008, ho1, Kan) -> No colonies on plates | ||
*#[[Melbourne/BBa_I15009|'''P2 21C''']] - (BBa_I15009, PcyA, Kan) ->No colonies on plates | *#[[Melbourne/BBa_I15009|'''P2 21C''']] - (BBa_I15009, PcyA, Kan) ->No colonies on plates | ||
| Line 10: | Line 10: | ||
| - | *26 June 2007: [[Melbourne/IGEM 2007 kit| | + | *26 June 2007: [[Melbourne/IGEM 2007 kit|Resuspended the following from Registry plates]] & [[Melb:Transformation Protocol|Transformed]] into Joe's competent DH5alpha cells. |
*#[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034, RBS, Amp) | *#[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034, RBS, Amp) | ||
*#[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051, c1 protein, Amp) | *#[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051, c1 protein, Amp) | ||
| Line 77: | Line 77: | ||
===30 June 2007=== | ===30 June 2007=== | ||
==Week 2== | ==Week 2== | ||
| - | + | *2 July 2007: [[Melbourne/IGEM 2007 kit|Resuspended the following from Registry plates]] & [[Melb:Transformation Protocol|Transformed]] into Joe's competent DH5alpha cells. | |
| - | + | *#Q04510 | |
| - | + | *#E0241 | |
| + | *#E0040 | ||
| + | *#B0014 | ||
| + | *#J61035 | ||
| + | |||
| + | *3 July 2007: [[Melb:Transformation Protocol|Re Transformed]] into Joe's competent DH5alpha cells. Each Plate had additional 40uL of 50mg/ml Ampicillan spread on surface. | ||
| + | *#[[Melbourne/BBa_J61035|'''P4 8J''']] -> Three colonies -> grew in liquid culture 4 july | ||
| + | *#[[Melbourne/BBa_E0040|'''P1 5H''']] -> Multiple colonies at edge -> did not grow in overnight liquid culture 4 july 'poor amp spreading' | ||
| + | *#[[Melbourne/BBa_E0241|'''P2 15L''']] -> Three colonies -> grew in liquid culture 4 july | ||
| + | |||
| + | *4 July 2007: | ||
====Ampicillin Plates==== | ====Ampicillin Plates==== | ||
Revision as of 07:53, 5 July 2007
Contents |
Week 1
- 25 June 2007: Prepared LB agar plates Amp & Kana.
- 25 June 2007: Resuspended the following from Registry plates & Transformed (shorter protocol) into competent DH5alpha cells.
- 26 June 2007: Resuspended the following from Registry plates & Transformed into Joe's competent DH5alpha cells.
- 26 June 2007: Streaked the following cells:
- pJS010 (from solid agar, Amp)
- Fusion protein (from glycerol stock, Amp?)
- BBa_I15010 (from solid agar, Kan)
- 27 June 2007: Transformed into Joe's competent DH5alpha cells.
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: Resuspended the following from Registry plates & Transformed into Joe's competent DH5alpha cells.
- Q04510
- E0241
- E0040
- B0014
- J61035
- 3 July 2007: Re Transformed into Joe's competent DH5alpha cells. Each Plate had additional 40uL of 50mg/ml Ampicillan spread on surface.
- 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)
