Melbourne/Lab Notebook

From 2007.igem.org

< Melbourne(Difference between revisions)
(→Miniprep)
 
(43 intermediate revisions not shown)
Line 1: Line 1:
[[Melbourne|<Back to team home page>]]  
[[Melbourne|<Back to team home page>]]  
 +
*Gas Vesicle lab notebook:
 +
** [[Melbourne/Lab GV Notebook|Gas Vesicles Notebook]]
-
==Week 1==
+
*Blue photoreceptor notebook:
-
===25 June 2007===
+
** [[Melbourne/Lab BL Notebook|Blue Photoreceptor Notebook]]
-
Prepared LB agar plates.
+
*Photoreceptor & control system lab notebooks:
-
 
+
** [[Melbourne/Lab Notebook Weeks 1-4|Weeks 1-4]]
-
====Transformation====
+
** [[Melbourne/Lab Notebook Weeks 5-8|Weeks 5-8]]
-
#resuspended the following from Registry plates:
+
** [[Melbourne/Lab Notebook Weeks 9-12|Weeks 9-12]]
-
#*[[Melbourne/BBa_I15008|'''P2 21A''']] - (BBa_I15008, ho1, Kan)
+
** [[Melbourne/Lab Notebook Weeks 13-16|Weeks 13-16]]
-
#*[[Melbourne/BBa_I15009|'''P2 21C''']] - (BBa_I15009, PcyA, Kan)
+
** [[Melbourne/Lab Notebook Weeks 17-18|Weeks 17-18]]
-
#*[[Melbourne/BBa_R0084|'''P1 11H''']] - (BBa_R0084, OmpR positive promoter, Amp)
+
-
#Punctured foil with pipette tip.
+
-
#Resuspended in 15uL ddH2O.
+
-
#Stored in-20 (after taking 1uL for transformation).
+
-
#[[Melb:Transformation Protocol|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:
+
-
#*[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034, RBS, Amp)
+
-
#*[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051, c1 protein, Amp)
+
-
#*[[Melbourne/BBa_B0010|'''P2 3P''']] (BBa_B0010, Terminator, Amp)
+
-
*Think some DNA may have remained in wells
+
-
#[[Melb:Transformation Protocol|Transformed]] into competent DH5alpha cells from Joe
+
-
 
+
-
====Streak plates====
+
-
#Streaked the following cells:
+
-
#*[[Melbourne/pJS010|'''pJS010''']] (from solid agar, Amp)
+
-
#*[[Melbourne/Fusion|'''Fusion protein''']] (from glycerol stock, Amp?)
+
-
#*[[Melbourne/BBa_I15010|'''BBa_I15010''']] (from solid agar, Kan)
+
-
 
+
-
===27 June 2007===
+
-
====Transformation====
+
-
#Repeated transformation of failed parts from Monday:
+
-
##[[Melbourne/BBa_I15008|'''P2 21A''']] (Kan)
+
-
##[[Melbourne/BBa_I15009|'''P2 21C''']] (Kan)
+
-
*Used resuspended DNA that was stored on Monday
+
-
 
+
-
====Liquid culture====
+
-
#[[Melb:Growing up cells|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:
+
-
#*[[Melbourne/pJS010|'''pJS010''']]
+
-
#*[[Melbourne/Fusion|'''Fusion''']]
+
-
#*[[Melbourne/BBa_B0034|'''P1 3O''']]
+
-
#*[[Melbourne/BBa_C0051|'''P1 5G''']]
+
-
#*[[Melbourne/BBa_B0010|'''P2 3P''']]
+
-
#*[[Melbourne/BBa_R0084|'''P1 11H''']]
+
-
#To the Kan LB a single colony from the transformation plate of [[Melbourne/BBa_I15010|'''BBa_I15010''']] was introduced.
+
-
#Cells incubated at 37degrees with shaking overnight.
+
-
 
+
-
===28 June 2007===
+
-
 
+
-
====Miniprep====
+
-
#[[Melb:Miniprep protocol|Minipreped]] the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
+
-
#*[[Melbourne/pJS010|'''pJS010''']]
+
-
#*[[Melbourne/Fusion|'''Fusion''']]
+
-
#*[[Melbourne/BBa_B0034|'''P1 3O''']]
+
-
#*[[Melbourne/BBa_C0051|'''P1 5G''']]
+
-
#*[[Melbourne/BBa_B0010|'''P2 3P''']]
+
-
#*[[Melbourne/BBa_R0084|'''P1 11H''']]
+
-
#*[[Melbourne/BBa_I15010|'''I15010''']]
+
-
#Stored in -20 freezer
+
-
 
+
-
====Digest====
+
-
 
+
-
 
+
-
====Liquid culture====
+
-
#[[Melb:Growing up cells|Cultured]] the following cells from transformed plates:
+
-
#*[[Melbourne/BBa_R0084|'''P1 11H''']] (BBa_R0084)
+
-
#*[[Melbourne/BBa_I15010|'''BBa_I15010''']]
+
-
#*[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034)
+
-
#*[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051)
+
-
#*[[Melbourne/BBa_I15008|'''P1 21A''']] (BBa_I15008)
+
-
#*[[Melbourne/BBa_I15009|'''P1 21C''']] (BBa_I15009)
+
-
 
+
-
===29 June 2007===
+
-
====Miniprep====
+
-
#[[Melb:Miniprep protocol|Minipreped]] the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
+
-
#*[[Melbourne/BBa_R0084|'''P1 11H''']] (BBa_R0084)
+
-
#*[[Melbourne/BBa_I15010|'''BBa_I15010''']]
+
-
#*[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034)
+
-
#*[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051)
+
-
#*[[Melbourne/BBa_I15008|'''P1 21A''']] (BBa_I15008)
+
-
#*[[Melbourne/BBa_I15009|'''P1 21C''']] (BBa_I15009)
+
-
 
+
-
#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====
+
-
*[[Melbourne/Growing up cells|Cultured]] 2 colonies from each of the following transformed plates and labelled as follows
+
-
**[[Melbourne/BBa_B0010|'''P2 3P 1''']]
+
-
**[[Melbourne/BBa_B0010|'''P2 3P 2''']]
+
-
**[[Melbourne/BBa_I15010|'''I15010 1''']]
+
-
**[[Melbourne/BBa_I15010|'''I15010 2''']]
+
-
**[[Melbourne/pJS010|'''pJS010 1''']]
+
-
**[[Melbourne/pJS010|'''pJS010 2''']]
+
-
**[[Melbourne/Fusion|'''Fusion 1''']]
+
-
**[[Melbourne/Fusion|'''Fusion 2''']]
+
-
**[[Melbourne/BBa_J61035|'''P4 8J 1''']]
+
-
**[[Melbourne/BBa_J61035|'''P4 8J 2''']]
+
-
**[[Melbourne/BBa_E0241|'''P2 15L 1''']]
+
-
**[[Melbourne/BBa_E0241|'''P2 15L 2''']]
+
-
**[[Melbourne/BBa_Q04510|'''P2 13K 1''']]
+
-
**[[Melbourne/BBa_Q04510|'''P2 13K 2''']]
+
-
**[[Melbourne/BBa_B0014|'''P1 1G 1''']]
+
-
**[[Melbourne/BBa_B0014|'''P1 1G 2''']]
+
-
**[[Melbourne/BBa_R0084|'''P1 11H 1''']]
+
-
**[[Melbourne/BBa_R0084|'''P1 11H 2''']]
+
-
**[[Melbourne/BBa_E0040|'''P1 5H 1''']]
+
-
**[[Melbourne/BBa_E0040|'''P1 5H 2''']]
+
-
**suspected contaminating colony from [[Melbourne/BBa_J61035|'''P4 8J''']]
+
-
 
+
-
===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.
+
-
**[[Melbourne/BBa_B0010|'''P2 3P 1''']]
+
-
**[[Melbourne/BBa_B0010|'''P2 3P 2''']]
+
-
**[[Melbourne/BBa_I15010|'''I15010 1''']]
+
-
**[[Melbourne/BBa_I15010|'''I15010 2''']]
+
-
**[[Melbourne/pJS010|'''pJS010 1''']]
+
-
**[[Melbourne/pJS010|'''pJS010 2''']]
+
-
**[[Melbourne/Fusion|'''Fusion 1''']]
+
-
**[[Melbourne/Fusion|'''Fusion 2''']]
+
-
**[[Melbourne/BBa_J61035|'''P4 8J 1''']]
+
-
**[[Melbourne/BBa_J61035|'''P4 8J 2''']]
+
-
**[[Melbourne/BBa_E0241|'''P2 15L 1''']]
+
-
**[[Melbourne/BBa_E0241|'''P2 15L 2''']]
+
-
**[[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_B0014|'''P1 1G 1''']]
+
-
**[[Melbourne/BBa_B0014|'''P1 1G 2''']]
+
-
*the following liquid cultures were not miniprepped due to failure (no growth)
+
-
**[[Melbourne/BBa_R0084|'''P1 11H 1''']]
+
-
**[[Melbourne/BBa_R0084|'''P1 11H 2''']]
+
-
**[[Melbourne/BBa_E0040|'''P1 5H 1''']]
+
-
**[[Melbourne/BBa_E0040|'''P1 5H 2''']]
+
-
**suspected contaminating colony from [[Melbourne/BBa_J61035|'''P4 8J''']]
+
-
 
+
-
===6 July 2007===
+
-
==Week 3==
+
-
===9 July 2007===
+
-
===10 July 2007===
+
-
===11 July 2007===
+
-
===12 July 2007===
+
-
===13 July 2007===
+
-
==Week 4==
+
-
===16 July 2007===
+
-
===17 July 2007===
+
-
===18 July 2007===
+
-
===19 July 2007===
+
-
===20 July 2007===
+
-
==Week 5==
+
-
===23 July 2007===
+
-
===24 July 2007===
+
-
===25 July 2007===
+
-
===26 July 2007===
+
-
===27 July 2007===
+
-
==Week 6==
+
-
===30 July 2007===
+
-
===31 July 2007===
+
-
===1  Aug 2007===
+
-
===2  Aug 2007===
+
-
===3  Aug 2007===
+

Latest revision as of 14:25, 26 October 2007

<Back to team home page>

Retrieved from "http://2007.igem.org/wiki/index.php/Melbourne/Lab_Notebook"