Team:Groningen/Notebook/17 July 2009

From 2009.igem.org

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[[Image:PCR15-7-19.jpg]] [[Image:F102471_2009-07-17_05hr_30min_A_E,_small_band.jpg]]
[[Image:PCR15-7-19.jpg]] [[Image:F102471_2009-07-17_05hr_30min_A_E,_small_band.jpg]]
-
Because of the negative results we will try to get positive results with an other polymerase enzyme; Phusion. And try the cloning again as planned. PCR1 Fw,mut1RC should give product of 1153bp and PCR2 Rev,mut2RC a 261bp size product. Further more we will test a PCR with pBAD 24 Fw,rev to get smaller templates (2222 bp).
+
Because of the negative results we will try to get positive results with an other polymerase enzyme; Phusion. And try the cloning again as planned. PCR1 Fw,mut1RC should give product of 1153bp and PCR2 Rev,mut2RC a 261bp size product. Further more we will test a PCR with pBAD 24 Fw,rev to test the template and look at options to get smaller templates (2222 bp).
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Revision as of 15:00, 17 July 2009

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


Wet

GVP Cluster

Discussion:

All colonies (transformants vector + promoter ligated with gvp) and o/n culture became red, so probably RFP is still in there... What might be the problem? The vector with promoter self-ligated due to uncomplete digestion (done 15 July 2009) leading to the following fragments:

  • vector+RFP linear
  • vector
  • RFP

When the upper fragment (~2kb) is contaminated with linear vector this would lead high efficiency self-ligation instead of ligating gvp with the emtpy vector.

TODO Therefore the restriction/purification/ligation will be redone and selection of non-red colonies will be made!
DONE Also plasmid from the o/n cultures will be purified and analysed by restriction analysis.
  • For plasmid isolation the Sigma Plasmid isolation kit was used (eluted in 50ul MQ).
  • DNA concentrations:
Sample ng/ul 260/280 260/230
gpv-promoter test (colony 9) 82.5 2.02 2.21
gpv-promoter test (colony 10) 71.9 1.96 2.21
  • Restriction analysis with SpeI and PstI to find insert length.
    • Expected length of with / with RFP insert: 2096bp(promoter + plasmid backbone), 887bp(RFP)
    • Expected length of with gvp insert: 2096bp(promoter + plasmid backbone),6096bp(gvp)
    • Expected length of with gpv insert: 3513(plasmid backbone), 6096 (GVP cluster)

Restriction mixture for J23100+gvp and GVP

  • 10 μL promoter plasmid (61.4 ng/μL, 96.1 ng/μL)
  • 6μL MQ
  • 2μL Fast digest buffer
  • 1μL PstI fast digest enzyme
  • 1μL SpeI fast digest enzyme

10ul was loaded on a 1%TBE agarose gel and ran for 30 @ 100V (V const)

2009-07-17 test gvp+prom-2.PNG Generulers 1kb marker Fermentas.jpg

→From left to right: 1kb marker, gvp-promoter () construct nr 9, nr 10, gvp in , promoter ()

So the assumption that the vector with the promoter self-ligated was right, as the gvp-promoter construct has the same bands as the promoter has and the size of the bands are as expected for a RFP insert.

Transporters

Below yesterdays PCRs, There seems to be a vague band at ~1000 in A1 and E1. therefore loaded the last 20 uL PCR Product on gel as well. showing an expected product op 970bp.

PCR15-7-19.jpg F102471 2009-07-17 05hr 30min A E, small band.jpg

Because of the negative results we will try to get positive results with an other polymerase enzyme; Phusion. And try the cloning again as planned. PCR1 Fw,mut1RC should give product of 1153bp and PCR2 Rev,mut2RC a 261bp size product. Further more we will test a PCR with pBAD 24 Fw,rev to test the template and look at options to get smaller templates (2222 bp).

PCR1
Component amount
2x Phusion MM 12.5 uL
MQ 9.5 uL
Fw 1 uL
mut1RC 1 uL
DNA 1 uL
PCR1 program Temperature Time
Denaturing 98° 1.30 min
Start Cycles 30X
Denaturing 98° 10 sec
Annealing 55° 20 sec
Elongation 72° 2:10 min
End cycles
Final elongation 72° 10 min
Hold Forever


PCR2
Component amount
2x Phusion MM 12.5 uL
MQ 9.5 uL
Rev 1 uL
mut2RC 1 uL
DNA 1 uL
PCR2 program Temperature Time
Denaturing 98° 1.30 min
Start Cycles 30X
Denaturing 98° 10 sec
Annealing 55° 20 sec
Elongation 72° 20 sec
End cycles
Final elongation 72° 10 min
Hold Forever


pBAD24
Component amount
mm NHSO 21 uL
24_F 1 uL
24_R 1 uL
DNA 1 uL
taq polymerase 1 uL
PCR pBAD-24 program Temperature Time
Denaturing 95° 2.00 min
Start Cycles 25X
Denaturing 95° 30 sec
Annealing 55° 20 sec
Elongation 72° 2.10 min
End cycles
Final elongation 72° 10 min
Hold Forever

Metal Accumulation

Vectors

Dry

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]