From 2009.igem.org
Wet
GVP Cluster
Restriction for Assembly
The vector ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J61035 BBa_J61035]) containing [http://partsregistry.org/wiki/index.php/Part:BBa_I750016 GVP] cluster was cut with PstI and XbaI to create correct ends for insert in promotor vector ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 BBa_J61002]).
- 10μL MQ
- 6μL plasmid in MQ (2μg) (329.7 ng/μL)
- 2μL Fast digest buffer
- 1μL PstI fast digest enzyme
- 1μL XbaI fast digest enzyme
The vector ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 BBa_J61002]) containing promotor [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] was cut with SpeI and PstI.
- 4μL MQ
- 12μL plasmid in MQ (155.4 ng/μL)
- 2μL Fast digest buffer
- 1μL SpeI fast digest enzyme
- 1μL PstI fast digest enzyme
The cups were incubated in a heat block at 37°C for 30 min. followed by adding 4μL 6x loading buffer for gel electroforesis.
Gel electroforesis
24μL of each sample was loaded on a 1% agarose gel (devided over two slots) with EtBr and a 1kb ladder was used (see picture).
- → From left to right: Empty slot, 1kb Marker, Empty slot, HmtA PCR reaction, Empty slot, (2x) GVP, Empty slot, (2x) [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100], and Empty slot
Gel Purification of GVP
"High Pure PCR Product Purification Kit" from [http://www.roche-applied-science.com Roche] was used for gel purification.
- ~200mg of gel containing the desired fragment was dissolved in 500μL binding buffer by heating to 60°C for 10 min. and carefully vortexing.
- 250μL isopropanol was added to the tube and vortexed
- dissolved gel was transfered to the column and centrifuged for 1 min.
- column was washed with 500μL Wash Solution, centrifuged for 1 min., 200μL Wash solution was added and centrifuged for 1 min.
- 15μL MQ was applied to the column, incubated for 3 min. at room temperature and centrifuged for 1 min. at full speed.
Concentration of GVP-vector
The concentration of isolated cut vector of GVP with EcoRI and XbaI was determined with the use of a nano-drop.
GVP-cluster eluted in MQ
- 33.5 ng/μL
- 1.65 (260/280)
- 0.77 (260/230)
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] + vector eluted in MQ
- 14.2 ng/μL
- 1.78 (260/280)
- 1.01 (260/230)
- → Concentrations are just high enough for a ligation reaction to be performed, some additional tuning of the procedure is required!!
Ligation of [http://partsregistry.org/wiki/index.php/Part:BBa_I750016 GVP] into [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 vector]
Mix:
- 1 μL T4 Ligase buffer
- 1 μL Vector (J23100 restriction fragment, 14.2 ng/μL)
- 7.6 μL Insert (GVP gene cluster, 33.5 ng/μL)
- 0.5 μL T4 Ligase
Method:
- The mixture is incubated 30 min, 26°C
- To inactivate Ligase 10 min 70°C
Transformation of E.coli TOP10 cells with GVP + J23100
- 2 μL Ligation mix added to cells from -80°C glycerol stock
- 30 min incubation on ice
- 5 min heat shock 37°C
- 5 min on ice
- Cells added to 3 ml Ty in a reaction tube
- Incubation 1 h in waterbath shaker 37°C
- The cells are plated on Ty agar (100 μg/ml)
- On one plate 200 μL is streaked out. The rest is concentrated (centrifugation: 1 min, 14600rpm) in 200 μL and also plated.
- Incubation o/n 37°C
Transporters
We decided to make MasterMixes of our own. All but 1uL template, 2uL primers and 1uL taq for 25uL reactions.
1X MasterMix Buffer NH SO
Component
| concentrations
| volumes
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dNTP
| 0.2 mM
| 8 uL
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MgCl
| 2.0 mM
| 80 uL
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taq buffer KCl
| 1X
| 100 uL
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MQ
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| 652 uL
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1X MasterMix Buffer KCl
Component
| concentrations
| volumes
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dNTP
| 0.2 mM
| 8 uL
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MgCl
| 1.5 mM
| 60 uL
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taq buffer (NH)SO
| 1X
| 100 uL
|
MQ
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| 672 uL
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MasterMix3
Component | Mg/K
|
MQ
| 28.5 uL
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F mut1
| 2 uL
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R mut2
| 2 uL
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dNTP
| 2 uL
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MgCl
| 3 uL
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taq buffer KCl
| 5 uL
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taq buffer (NH)SO
| 5 uL
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taq polymerase
| 2.5 uL
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PCR 3 program
| Temperature
| Time
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Denaturing
| 95°
| 2.00 min
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| Start Cycles 25X
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Denaturing
| 95°
| 30 sec
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Annealing
| 55°
| 20 sec
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Elongation
| 72°
| 2.10 min
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| End cycles
|
Final elongation
| 72°
| 10 min
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Hold
| 4°
| Forever
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Vectors
- overnight cultures were diluted 100, 200 or 400 x
- 200 ul of culture per well
- Measured for 16 h
- - every 15 min
- - @ 37° with shaking 6 mm
- - for mRFP1 excitation =590 nm
- excitation =610 nm
96-wells Microtiter plate
| 1 LB
| 2 LB
| 3 LB
| 4 LB
| 5 LB
| 6
| 7 TY
| 8 TY
| 9 TY
| 10 TY
| 11 TY
| 12
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A 100x
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
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B 100x
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
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C 200x
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
|
D 200x
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
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E 400x
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
|
F 400x
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
|
| pSB1AC3
| pSB3K3
| BBA_J23109
| BBA_J23106
| BBA_J23100
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H
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Dry
April
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M | T | W | T | F | S | S
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| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 1]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 2]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 5]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 6]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 7]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 8]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 10]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 14]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 15]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 16]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/17_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 17]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 18]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 19]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 20]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 21]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 22]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 23]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 24]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 25]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 28]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 29]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
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May
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M | T | W | T | F | S | S
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| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 1]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 2]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
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[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 5]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 6]
| [http://2009.igem.org/Team:Groningen/Notebook/7_May_2009 7]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 8]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 10]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 14]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 15]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 16]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/17_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 17]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 18]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 19]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 20]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 21]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 22]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 23]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 24]
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[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 25]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
| [http://2009.igem.org/Team:Groningen/Notebook/28_May_2009 28]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 29]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/31_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 31]
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June
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M | T | W | T | F | S | S
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[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 1]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 2]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
| [http://2009.igem.org/Team:Groningen/Notebook/5_June_2009 5]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 6]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 7]
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[http://2009.igem.org/Team:Groningen/Notebook/8_June_2009 8]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 10]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 14]
|
[http://2009.igem.org/Team:Groningen/Notebook/15_June_2009 15]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 16]
| [http://2009.igem.org/Team:Groningen/Notebook/17_June_2009 17]
| [http://2009.igem.org/Team:Groningen/Notebook/18_June_2009 18]
| [http://2009.igem.org/Team:Groningen/Notebook/19_June_2009 19]
| [http://2009.igem.org/Team:Groningen/Notebook/20_June_2009 20]
| [http://2009.igem.org/Team:Groningen/Notebook/21_June_2009 21]
|
[http://2009.igem.org/Team:Groningen/Notebook/22_June_2009 22]
| [http://2009.igem.org/Team:Groningen/Notebook/23_June_2009 23]
| [http://2009.igem.org/Team:Groningen/Notebook/24_June_2009 24]
| [http://2009.igem.org/Team:Groningen/Notebook/25_June_2009 25]
| [http://2009.igem.org/Team:Groningen/Notebook/26_June_2009 26]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 28]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 29]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
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July
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M | T | W | T | F | S | S
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| [http://2009.igem.org/Team:Groningen/Notebook/1_July_2009 1]
| [http://2009.igem.org/Team:Groningen/Notebook/2_July_2009 2]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 5]
|
[http://2009.igem.org/Team:Groningen/Notebook/6_July_2009 6]
| [http://2009.igem.org/Team:Groningen/Notebook/7_July_2009 7]
| [http://2009.igem.org/Team:Groningen/Notebook/8_July_2009 8]
| [http://2009.igem.org/Team:Groningen/Notebook/9_July_2009 9]
| [http://2009.igem.org/Team:Groningen/Notebook/10_July_2009 10]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
|
[http://2009.igem.org/Team:Groningen/Notebook/13_July_2009 13]
| [http://2009.igem.org/Team:Groningen/Notebook/14_July_2009 14]
| [http://2009.igem.org/Team:Groningen/Notebook/15_July_2009 15]
| [http://2009.igem.org/Team:Groningen/Notebook/16_July_2009 16]
| [http://2009.igem.org/Team:Groningen/Notebook/17_July_2009 17]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 18]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 19]
|
[http://2009.igem.org/Team:Groningen/Notebook/20_July_2009 20]
| [http://2009.igem.org/Team:Groningen/Notebook/21_July_2009 21]
| [http://2009.igem.org/Team:Groningen/Notebook/22_July_2009 22]
| [http://2009.igem.org/Team:Groningen/Notebook/23_July_2009 23]
| [http://2009.igem.org/Team:Groningen/Notebook/24_July_2009 24]
| [http://2009.igem.org/Team:Groningen/Notebook/25_July_2009 25]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
|
[http://2009.igem.org/Team:Groningen/Notebook/27_July_2009 27]
| [http://2009.igem.org/Team:Groningen/Notebook/28_July_2009 28]
| [http://2009.igem.org/Team:Groningen/Notebook/29_July_2009 29]
| [http://2009.igem.org/Team:Groningen/Notebook/30_July_2009 30]
| [http://2009.igem.org/Team:Groningen/Notebook/31_July_2009 31]
|
|
August
|
M | T | W | T | F | S | S
|
|
|
|
|
| [http://2009.igem.org/Team:Groningen/Notebook/1_August_2009 1]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_August_2009&preload=Team:Groningen/Notebook/Day&action=edit 2]
|
[http://2009.igem.org/Team:Groningen/Notebook/3_August_2009 3]
| [http://2009.igem.org/Team:Groningen/Notebook/4_August_2009 4]
| [http://2009.igem.org/Team:Groningen/Notebook/5_August_2009 5]
| [http://2009.igem.org/Team:Groningen/Notebook/6_August_2009 6]
| [http://2009.igem.org/Team:Groningen/Notebook/7_August_2009 7]
| [http://2009.igem.org/Team:Groningen/Notebook/8_August_2009 8]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_August_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
|
[http://2009.igem.org/Team:Groningen/Notebook/10_August_2009 10]
| [http://2009.igem.org/Team:Groningen/Notebook/11_August_2009 11]
| [http://2009.igem.org/Team:Groningen/Notebook/12_August_2009 12]
| [http://2009.igem.org/Team:Groningen/Notebook/13_August_2009 13]
| [http://2009.igem.org/Team:Groningen/Notebook/14_August_2009 14]
| [http://2009.igem.org/Team:Groningen/Notebook/15_August_2009 15]
| [http://2009.igem.org/Team:Groningen/Notebook/16_August_2009 16]
|
[http://2009.igem.org/Team:Groningen/Notebook/17_August_2009 17]
| [http://2009.igem.org/Team:Groningen/Notebook/18_August_2009 18]
| [http://2009.igem.org/Team:Groningen/Notebook/19_August_2009 19]
| [http://2009.igem.org/Team:Groningen/Notebook/20_August_2009 20]
| [http://2009.igem.org/Team:Groningen/Notebook/21_August_2009 21]
| [http://2009.igem.org/Team:Groningen/Notebook/22_August_2009 22]
| [http://2009.igem.org/Team:Groningen/Notebook/23_August_2009 23]
|
[http://2009.igem.org/Team:Groningen/Notebook/24_August_2009 24]
| [http://2009.igem.org/Team:Groningen/Notebook/25_August_2009 25]
| [http://2009.igem.org/Team:Groningen/Notebook/26_August_2009 26]
| [http://2009.igem.org/Team:Groningen/Notebook/27_August_2009 27]
| [http://2009.igem.org/Team:Groningen/Notebook/28_August_2009 28]
| [http://2009.igem.org/Team:Groningen/Notebook/29_August_2009 29]
| [http://2009.igem.org/Team:Groningen/Notebook/30_August_2009 30]
|
[http://2009.igem.org/Team:Groningen/Notebook/31_August_2009 31]
|
|
September
|
M | T | W | T | F | S | S
|
| [http://2009.igem.org/Team:Groningen/Notebook/1_September_2009 1]
| [http://2009.igem.org/Team:Groningen/Notebook/2_September_2009 2]
| [http://2009.igem.org/Team:Groningen/Notebook/3_September_2009 3]
| [http://2009.igem.org/Team:Groningen/Notebook/4_September_2009 4]
| [http://2009.igem.org/Team:Groningen/Notebook/5_September_2009 5]
| [http://2009.igem.org/Team:Groningen/Notebook/6_September_2009 6]
|
[http://2009.igem.org/Team:Groningen/Notebook/7_September_2009 7]
| [http://2009.igem.org/Team:Groningen/Notebook/8_September_2009 8]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
| [http://2009.igem.org/Team:Groningen/Notebook/10_September_2009 10]
| [http://2009.igem.org/Team:Groningen/Notebook/11_September_2009 11]
| [http://2009.igem.org/Team:Groningen/Notebook/12_September_2009 12]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
|
[http://2009.igem.org/Team:Groningen/Notebook/14_September_2009 14]
| [http://2009.igem.org/Team:Groningen/Notebook/15_September_2009 15]
| [http://2009.igem.org/Team:Groningen/Notebook/16_September_2009 16]
| [http://2009.igem.org/Team:Groningen/Notebook/17_September_2009 17]
| [http://2009.igem.org/Team:Groningen/Notebook/18_September_2009 18]
| [http://2009.igem.org/Team:Groningen/Notebook/19_September_2009 19]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 20]
|
[http://2009.igem.org/Team:Groningen/Notebook/21_September_2009 21]
| [http://2009.igem.org/Team:Groningen/Notebook/22_September_2009 22]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 23]
| [http://2009.igem.org/Team:Groningen/Notebook/24_September_2009 24]
| [http://2009.igem.org/Team:Groningen/Notebook/25_September_2009 25]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
|
[http://2009.igem.org/Team:Groningen/Notebook/28_September_2009 28]
| [http://2009.igem.org/Team:Groningen/Notebook/29_September_2009 29]
| [http://2009.igem.org/Team:Groningen/Notebook/30_September_2009 30]
|
|
October
|
M | T | W | T | F | S | S
|
|
|
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 1]
| [http://2009.igem.org/Team:Groningen/Notebook/2_October_2009 2]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
|
[http://2009.igem.org/Team:Groningen/Notebook/5_October_2009 5]
| [http://2009.igem.org/Team:Groningen/Notebook/6_October_2009 6]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 7]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 8]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 10]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 14]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 15]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 16]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/17_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 17]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 18]
|
[http://2009.igem.org/Team:Groningen/Notebook/19_October_2009 19]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 20]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 21]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 22]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 23]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 24]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 25]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 28]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 29]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/31_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 31]
|
|
November
|
M | T | W | T | F | S | S
|
|
|
|
|
|
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 1]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 2]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 5]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 6]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 7]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 8]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 10]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
| [http://2009.igem.org/Team:Groningen/Notebook/14_November_2009 14]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 15]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 16]
| [http://2009.igem.org/Team:Groningen/Notebook/17_November_2009 17]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 18]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 19]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 20]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 21]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 22]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 23]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 24]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 25]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 28]
| [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 29]
|
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
|
|