Melbourne/Lab Notebook
From 2007.igem.org
(Difference between revisions)
(→27 June 2007) |
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#Streaked the following cells: | #Streaked the following cells: | ||
*PJS010 (from solid agar, Amp) | *PJS010 (from solid agar, Amp) | ||
| - | *Fusion protein (from glycerol stock, | + | *Fusion protein (from glycerol stock, Amp?) |
*BBa_I15010 (from solid agar, Kan) | *BBa_I15010 (from solid agar, Kan) | ||
Revision as of 01:44, 3 July 2007
Contents |
Week 1
25 June 2007
Prepared LB agar plates.
Transformation
- resuspended the following from Registry plates:
- BBa_I15008 - ho1 (P2 21A, Kan)
- BBa_I15009 - PcyA (P2 21C, Kan)
- BBa_R0084 - OmpR positive promoter (P1 11H, Amp)
- 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:
- BBa_B0034 (RBS, plate 1 3O, Amp)
- BBa_C0051 (C1 repressor, plate 1 5G, Amp)
- BBa_B0010 (Terminator, plate 2 3P, Amp)
- 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
- Repeated transformation of failed parts from Monday:
- BBa_I15008 (Kan)
- BBa_I15009 (Kan)
- 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 the following were introduced:
