Melbourne/Lab Notebook

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[[Melbourne|<Back to team home page>]]  
[[Melbourne|<Back to team home page>]]  
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==Week 1==
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*Gas Vesicle lab notebook:
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===25 June 2007===
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** [[Melbourne/Lab GV Notebook|Gas Vesicles Notebook]]
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Prepared LB agar plates.
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*Blue photoreceptor notebook:
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** [[Melbourne/Lab BL Notebook|Blue Photoreceptor Notebook]]
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====Transformation====
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*Photoreceptor & control system lab notebooks:
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#resuspended the following from Registry plates:
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** [[Melbourne/Lab Notebook Weeks 1-4|Weeks 1-4]]
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**BBa_I15008 - ho1 (P2 21A, Kan)
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** [[Melbourne/Lab Notebook Weeks 5-8|Weeks 5-8]]
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**BBa_I15009 - PcyA (P2 21C, Kan)
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** [[Melbourne/Lab Notebook Weeks 9-12|Weeks 9-12]]
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**BBa_R0084 - OmpR positive promoter (P1 11H, Amp)
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** [[Melbourne/Lab Notebook Weeks 13-16|Weeks 13-16]]
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##Punctured foil with pipette tip.
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** [[Melbourne/Lab Notebook Weeks 17-18|Weeks 17-18]]
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##Resuspended in 15uL ddH2O.
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##Stored in-20 (after taking 1uL for transformation).
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#[[Melb:Transformation Protocol|Transformed]] into competent DH5alpha cells with shorter incubation times as follows:
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**30min on ice after DNA addition
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**10min on ice after heat shock
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**30min at 37degrees with LB
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===26 June 2007===
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====Transformation from Monday====
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*Transformation of BBa_I15008 and BBa_I15009 failed.  No colonies on plates
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*Small number of colonies on BBa_R0084 plate.
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*Placed in cool room
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====Transformation====
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#Resuspended the following parts in 15uL:
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*BBa_B0034 (RBS, plate 1 3O, Amp)
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*BBa_C0051 (C1 repressor, plate 1 5G, Amp)
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*BBa_B0010 (Terminator, plate 2 3P, Amp)
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**Think some DNA may have remained in wells
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#[[Melb:Transformation Protocol|Transformed]] into competent DH5alpha cells from Joe
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====Streak plates====
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#Streaked the following cells:
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*PJS010 (from solid agar, Amp)
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*Fusion protein (from glycerol stock, Amp?)
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*BBa_I15010 (from solid agar, Kan)
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===27 June 2007===
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====Transformation====
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#Repeated transformation of failed parts from Monday:
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*BBa_I15008 (Kan)
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*BBa_I15009 (Kan)
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**Used resuspended DNA that was stored on Monday
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====Liquid culture====
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#[[Melb:Growing up cells|Prepared]] 30mL of Amp LB (100ug/mL). Prepared 5mL Kan LB (50mg/mL).
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#Aliquoted 5mL Amp LB into 6 50mL falcon tubes
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#To the Amp LB aliquots single transformed colonies from plates of the following were introduced:
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##PJS010
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##Fusion
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##BBa_B0034
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##BBa_C0051
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##BBa_B0010
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##BBa_R0084
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#To the Kan LB a single colony from the transformation plate of BBa_I15010 was introduced.
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#Cells incubated at 37degrees with shaking overnight.
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===28 June 2007===
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====Miniprep====
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#[[Melb:Miniprep protocol|Minipreped]] the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
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##PJS010
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##Fusion
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##BBa_B0034
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##BBa_C0051
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##BBa_B0010
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##BBa_R0084
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##BBa_I15010
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#Stored in -20 freezer
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====Digest====
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#
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===29 June 2007===
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===30 June 2007===
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==Week 2==
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===2 July 2007===
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=Now=
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===3 July 2007===
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===4 July 2007===
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===5 July 2007===
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===6 July 2007===
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==Week 3==
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===9 July 2007===
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===10 July 2007===
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===11 July 2007===
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===12 July 2007===
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===13 July 2007===
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==Week 4==
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===16 July 2007===
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===17 July 2007===
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===18 July 2007===
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===19 July 2007===
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===20 July 2007===
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==Week 5==
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===23 July 2007===
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===24 July 2007===
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===25 July 2007===
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===26 July 2007===
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===27 July 2007===
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==Week 6==
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===30 July 2007===
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===31 July 2007===
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===1  Aug 2007===
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===2  Aug 2007===
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===3  Aug 2007===
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Latest revision as of 14:25, 26 October 2007

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