Team:Groningen/Notebook/18 August 2009

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Sample MBP-ArsR (200) was isolated from red organims, indicating the presence of RFP or similar, this was however not engineered in this plasmid. Therefore a deviant restriction pattern was to be expected. It appears as if two different plasmids were present in this sample.
Sample MBP-ArsR (200) was isolated from red organims, indicating the presence of RFP or similar, this was however not engineered in this plasmid. Therefore a deviant restriction pattern was to be expected. It appears as if two different plasmids were present in this sample.
Sample MBP-ArsR (50) was transformed (1 uL) into TOP10 cells and plated out, 50 and 200 uL on LB-agar Amp<sub>100</sub>.
Sample MBP-ArsR (50) was transformed (1 uL) into TOP10 cells and plated out, 50 and 200 uL on LB-agar Amp<sub>100</sub>.
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 +
 +
*'''Continue with checking ''E. coli'' TOP10 transformants + pSB1AC3-mymT'''
 +
**Isolate plasmids of o/n culture.
 +
**Check  by restriction analysis:
 +
***Cut 500ng vector with either by:
 +
:: ''Eco''RI and ''Spe''I  --> expected size of fragments is: 213 + 3300
 +
:: or only by ''Pst''I  --> expected size of fragments is: 150 + 3300
 +
**Run both the colony PCR and the restriction product on a 1% Agarose gel.
 +
[[Image:F102471 2009-08-18 mymT pcr digest.jpg|350px]]
 +
**From left to right:(upper) 1kb marker, colony PCR on colonies 1-4 (MymT primers), colony PCR on colonies 1-4 (VR/VF2 primers); (lower) digest on 1-4 (EcoRI-SpeI), digest on 1-4 (PstI).
 +
**As there was no positive (with MymT insert) colony, the colony PCR was redone on 22 more colonies. This also didn't give any positive colony.
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*'''PCR to amplify fMT and MymT-RBS-pre'''
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**Use the plasmid pMAL-MT-GlpF (isolated 18/08/09) and PCR product MymT-RBS-pre/suff (14/08/09) to amplify the genes.
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**PCR was done using Phusion DNA polymerase, and the following primers:
 +
::fMT-RBS FW & fMT-suff RV
 +
::MymT RV & F1-RBS-ATG FW
 +
**PCR program with TouchDown 10x (Tm62-50&deg;C and Elongation time 30sec) and Cycles 30x (Tm 58&deg;C).
===Vectors===
===Vectors===

Revision as of 16:20, 21 August 2009

[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]

Wet

GVP Cluster

Plates

Showed single colony growth on plates with J61002-Metal Promoter-RFP plasmids, and were stored in the fridge for future preculture growth.

→ The plates with high and low concentration of transformed cells showed colonies in the expected ratio. On all plates 1 in 20 colonies was dark red, indication of ligation of the original high constitutive promoter back into the J61002 vector, or uncut plasmid which was transformed.

Over Night Cultures

→ All cultures showed growth, and can be used for plasmid isolation.
→ By growing colonies on medium with either amp. or kan. as a selection tool to find cultures with correct plasmid it was expected that only the tubes with kan. would show growth. Instead the transformed E.coli TOP10 cells seemed resistent to both antibiotics?
www.sigmaaldrich.com

Plasmid Purification

Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids [http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3] with [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 high], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 medium] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 low] constitutive promoters and GVP with the "Sygma-Aldrich™ [http://www.sigmaaldrich.com/life-science/molecular-biology/dna-and-rna-purification/plasmid-miniprep-kit.html GenElute™] Plasmid Miniprep Kit".

  • From each tube 4mL of culture was collected in a 2.0mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.
  • Plasmids were eluted with 30μL MQ and stored in the fridge


Concentration

Plasmid Conc. ng/μL 260/280 260/230 -20 box (michael Restriction Control
pSB3K3-BBa_J23100-GVP (amp.1) 39.0 1.82 0.95 No Negative
pSB3K3-BBa_J23100-GVP (amp.2) 23.5 2.11 1.51 No Negative
pSB3K3-BBa_J23100-GVP (kan.1) 20.3 2.52 2.72 No Negative
pSB3K3-BBa_J23100-GVP (kan.2) 23.0 2.17 2.56 No Negative
pSB3K3-BBa_J23109-GVP (amp.1) 29.1 2.05 1.69 No Negative
pSB3K3-BBa_J23109-GVP (amp.2) 40.6 1.92 1.16 No Negative
pSB3K3-BBa_J23109-GVP (kan.1) 44.1 1.88 1.08 No Negative
pSB3K3-BBa_J23109-GVP (kan.2) 24.6 2.26 2.22 No Negative


Restriction Control

Isolated plasmids were cut with fast-digest enzymes EcoRI and PstI to cut out GVP, and create ~6000bp and ~2700bp fragments.

Gel 18-8 no.1.jpg Generulers 1kb marker Fermentas.jpg

→ From left to right: (2x) 1kB ladder, (2x) pSB3K3-J23100-GVP (amp.), (2x) pSB3K3-J23100-GVP (kan.), (2x) pSB3K3-J23109-GVP (amp.), (2x) pSB3K3-J23109-GVP (kan.)
→ The restriction pattern was not expected (~6000bp and ~2700bp), and can be explained by ligation of vector pSB3K3 to J61035, which gives two EcoRI and one PstI site with ~2700bp, ~2000bp and ~1400bp fragments. The gel purification of GVP cluster must have gone wrong during the cutting of wanted fragment, and the vector instead of GVP has been isolated.
→ During growth of cells the negative control plate with ampicillin also contained colonies, and can be explained by both amp. and kan. resistence on the double vector ligation product.
→ New o.n. cultures for isolation of pSB3K3 plasmid with J23100, J23106, and J23109 promoters were made in 6mL LB-amp100 medium.

New Plates

From the plates with E.coli TOP10 containing the plasmid J61002 with three metal promoters infront of RFP, seven colonies of each were plated on a new LB-amp100 plate in a straight line to see if it are red or yellow colonies.

New Over Night Cultures

From each plate with E.coli TOP10 containing the plasmid J61002 with three metal promoters infront of RFP, four colonies were used in 5mL LB-amp100 medium for o.n. culture and isolation of plasmid tomorrow.

Transporters

HmtA curios about the dirty experiment. a 1% gel control will show if expected product is there. Unfortunately there was no product, the primer dimer changed and pcr 1 and 2 remain available. PCR again. Different program. Plus starting an other PCR1 and PCR2 to get new megaprimers.

Metal Accumulation

Restriction analysis MBP-ArsR

MBP-ArsR Fusion protein in pSB1AC3 restricted with XbaI & EcoRI - 18august2009.jpg

1% Agarose (1xTBE) gel with restriction analysis of MBP-ArsR fusion protein plasmids, with selfcloser (1), sample (white colony) (2), sample (red colony??) (3). Expected sizes of positive construct are shown. Samples were digested using [http://www.fermentas.com/catalog/re/fastpsti.htm PstI] & [http://www.fermentas.com/catalog/re/fastxbai.htm XbaI], ~45 min 37 °C. Sample MBP-ArsR (200) was isolated from red organims, indicating the presence of RFP or similar, this was however not engineered in this plasmid. Therefore a deviant restriction pattern was to be expected. It appears as if two different plasmids were present in this sample. Sample MBP-ArsR (50) was transformed (1 uL) into TOP10 cells and plated out, 50 and 200 uL on LB-agar Amp100.


  • Continue with checking E. coli TOP10 transformants + pSB1AC3-mymT
    • Isolate plasmids of o/n culture.
    • Check by restriction analysis:
      • Cut 500ng vector with either by:
EcoRI and SpeI --> expected size of fragments is: 213 + 3300
or only by PstI --> expected size of fragments is: 150 + 3300
    • Run both the colony PCR and the restriction product on a 1% Agarose gel.

F102471 2009-08-18 mymT pcr digest.jpg

    • From left to right:(upper) 1kb marker, colony PCR on colonies 1-4 (MymT primers), colony PCR on colonies 1-4 (VR/VF2 primers); (lower) digest on 1-4 (EcoRI-SpeI), digest on 1-4 (PstI).
    • As there was no positive (with MymT insert) colony, the colony PCR was redone on 22 more colonies. This also didn't give any positive colony.
  • PCR to amplify fMT and MymT-RBS-pre
    • Use the plasmid pMAL-MT-GlpF (isolated 18/08/09) and PCR product MymT-RBS-pre/suff (14/08/09) to amplify the genes.
    • PCR was done using Phusion DNA polymerase, and the following primers:
fMT-RBS FW & fMT-suff RV
MymT RV & F1-RBS-ATG FW
    • PCR program with TouchDown 10x (Tm62-50°C and Elongation time 30sec) and Cycles 30x (Tm 58°C).

Vectors

Restriction analysis metal promotors No promotors were found on gel (1% and 2%, see below).


Metal promotors in pSB1AC3 or pSB3K3 cut with SpeI & EcoRI - 18august2009.jpg

Restriction analysis of plasmids engineered to contain metal promotors. Constructs were digested using [http://www.fermentas.com/catalog/re/fastbcui.htm SpeI] & [http://www.fermentas.com/catalog/re/fastecori.htm EcoRI], expecting to cut out the ~100 bp that is the promotor. Gel is 2% agarose (1xTBE). DNA marker is 1 Kb.

Dry

Jasper worked on cleaning up the Mathematica notebooks for Meng2004 and Kostal2004, the newly derived rate constants for Meng2004 were put on the Wiki (on the transport page).


April
MTWTFSS
    [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]
May
MTWTFSS
        [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]
[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]
[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]
June
MTWTFSS
[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]
[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]
July
MTWTFSS
    [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
MTWTFSS
          [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
MTWTFSS
  [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
MTWTFSS
      [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
MTWTFSS
            [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]