Team:Newcastle/Labwork/27 July 2009

From 2009.igem.org

(Difference between revisions)
(New page: {{:Team:Newcastle/CSS}} {{:Team:Newcastle/Header}} {{:Team:Newcastle/Left}} Today we prepared the gel for last week's rescued plasmids whcih contain RFP and GFP biobricks. WE prepared the...)
 
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__NOTOC__
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[[Image:Team Newcastle 2009 iGEM ProbationaryP-Sign.PNG|50px|right]]
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=Formal Lab Session - 27th July 2009=
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[[Image:Team Newcastle 2009 iGEM 27-07-09 IMG 0178.JPG|250px|center]]
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Today we prepared the gel for last week's rescued plasmids whcih contain RFP and GFP biobricks. WE prepared the gel with 0.8% agorose. The final solution was 500ml using 4 gr of agorose in total.
 
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We checked the agar plates containing colonies of plasmids with five different biobricks.
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== Running the gel for RFP and GFP bioricks==
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Plate 1: Bba_C0056
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Today we prepared the gel with 0.8% agorose for last week's rescued plasmids which contain RFP and GFP biobricks. The final solution was 500ml using 4 gr of agorose in total.
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Plate 2: Bba_B1002
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Plate 3: Bba_
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Plate 4: Bba_C0076
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Plate 5: Bba_R0077
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Expcept from plate 3 we had colonies formed in all plates.
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We mixed 9ul of DNas with 1ul of the buffer and centrifuged shortly. The rest of the DNAs are then put back to the freezer.
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we prepared 12 tubes for 4 different biobricks. Colonies were picked and mixed in a tube containing 5ml of LB. Finally we left them at shaking incubator overnight.
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Finally we run the gel. We loaded the ladder to the first and fourth wells, GFP to the 2nd and RFP to the 3rd well and set the voltage to 120V which was then reduced to 90V.  
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We labelled the tubes as below:
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1A:
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1B:
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1C:
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We then prepared the plates for the plasmids.
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Below are the images:
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[[Image:Team Newcastle - iGem2009 - geldoc 2009-07-27.jpg|center]]
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 +
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==Transformation of new biobricks==
 +
We prepared the agar plates and left the tubes including the biobricks for incubation overnight.
 +
 +
We checked the agar plates containing colonies of plasmids with five different biobricks from last week. We will be following [https://2009.igem.org/Team:Newcastle/Project/Labwork/PhilsProtocols#Phil.E2.80.99s_mini_method_for_Alkaline_Lysis_for_Mini_Prep Phil’s mini method for Alkaline Lysis for Mini Prep] The list of plates and the biobricks are below:
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{| class="wikitable" border="1"
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|-
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| Plate
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| BioBrick
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| Number used for the labelling
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| Name
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|-
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| Plate 1
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| [http://partsregistry.org/Part:BBa_C0056 BBa_C0056]
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| 1
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| CI Coding Sequence
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|-
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| Plate 2
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| [http://partsregistry.org/Part:BBa_B1002 BBa_B1002]
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| 2
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| Terminator
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|-
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| Plate 3
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| [http://partsregistry.org/Part:BBa_C0077 BBa_C0077]
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| 3
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| CinR Coding sequence
 +
|-
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| Plate 4
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| [http://partsregistry.org/Part:BBa_C0076 BBa_C0076]
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| 4
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| CinI Coding Sequence
 +
|-
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| Plate 5
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| [http://partsregistry.org/Part:BBa_R0077 BBa_R0077]
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| 5
 +
| CinR sensitive promoter
 +
|-
 +
|}
 +
 +
Except from plate 3, we had colonies formed in all plates.
 +
 +
Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight.
 +
 +
==Preparation of the plates==
 +
[[Image:Team Newcastle 2009 iGEM 27-07-09 IMG 0177.JPG|200px|thumb|Goksel pouring out agar plates]]
 +
We prepared the agar solutions and poured into the plates which are then placed into the fridge.
 +
The list of plates and their content are as below.
 +
 +
{| class="wikitable" border="1"
 +
|-
 +
| Content
 +
| Number of plates prepared
 +
| Labelled as
 +
|-
 +
| LB
 +
| 4
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| LB 27/07/09
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|-
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| LB + Amp
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| 6
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| LB+A 27/07/09
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|-
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| LB + Amp + Kan
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| 5
 +
| LB+AK 27/07/09
 +
|-
 +
|}
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 +
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===Tomorrow===
 +
We have placed the required solutions to the bench for the experiment apart from RNAase. We need to find it first thing in the morning tomorrow then we will follow [https://2009.igem.org/Team:Newcastle/Project/Labwork/PhilsProtocols#Phil.E2.80.99s_mini_method_for_Alkaline_Lysis_for_Mini_Prep Phil’s mini method for Alkaline Lysis for Mini Prep]. GFP and RFP are placed into the freezer. The DNA ladder is in the fridge. Plates are placed into the fridge just below the top shelf. They were split into two since they did not all fit to the shelf.
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{{:Team:Newcastle/Project/Labwork/CalTemplate}}
{{:Team:Newcastle/Footer}}
{{:Team:Newcastle/Footer}}
{{:Team:Newcastle/Right}}
{{:Team:Newcastle/Right}}

Latest revision as of 17:21, 17 October 2009


Team Newcastle 2009 iGEM ProbationaryP-Sign.PNG

Formal Lab Session - 27th July 2009

Team Newcastle 2009 iGEM 27-07-09 IMG 0178.JPG


Running the gel for RFP and GFP bioricks

Today we prepared the gel with 0.8% agorose for last week's rescued plasmids which contain RFP and GFP biobricks. The final solution was 500ml using 4 gr of agorose in total.

We mixed 9ul of DNas with 1ul of the buffer and centrifuged shortly. The rest of the DNAs are then put back to the freezer.

Finally we run the gel. We loaded the ladder to the first and fourth wells, GFP to the 2nd and RFP to the 3rd well and set the voltage to 120V which was then reduced to 90V.

Below are the images:

Team Newcastle - iGem2009 - geldoc 2009-07-27.jpg


Transformation of new biobricks

We prepared the agar plates and left the tubes including the biobricks for incubation overnight.

We checked the agar plates containing colonies of plasmids with five different biobricks from last week. We will be following Phil’s mini method for Alkaline Lysis for Mini Prep The list of plates and the biobricks are below:

Plate BioBrick Number used for the labelling Name
Plate 1 [http://partsregistry.org/Part:BBa_C0056 BBa_C0056] 1 CI Coding Sequence
Plate 2 [http://partsregistry.org/Part:BBa_B1002 BBa_B1002] 2 Terminator
Plate 3 [http://partsregistry.org/Part:BBa_C0077 BBa_C0077] 3 CinR Coding sequence
Plate 4 [http://partsregistry.org/Part:BBa_C0076 BBa_C0076] 4 CinI Coding Sequence
Plate 5 [http://partsregistry.org/Part:BBa_R0077 BBa_R0077] 5 CinR sensitive promoter

Except from plate 3, we had colonies formed in all plates.

Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight.

Preparation of the plates

Goksel pouring out agar plates

We prepared the agar solutions and poured into the plates which are then placed into the fridge. The list of plates and their content are as below.

Content Number of plates prepared Labelled as
LB 4 LB 27/07/09
LB + Amp 6 LB+A 27/07/09
LB + Amp + Kan 5 LB+AK 27/07/09


Tomorrow

We have placed the required solutions to the bench for the experiment apart from RNAase. We need to find it first thing in the morning tomorrow then we will follow Phil’s mini method for Alkaline Lysis for Mini Prep. GFP and RFP are placed into the freezer. The DNA ladder is in the fridge. Plates are placed into the fridge just below the top shelf. They were split into two since they did not all fit to the shelf.

July
MTWTFSS
    [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/1_July_2009&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/2_July_2009&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/3_July_2009&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/4_July_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/5_July_2009&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/6_July_2009&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/7_July_2009&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/8_July_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/9_July_2009&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/10_July_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/11_July_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/12_July_2009&action=edit 12]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/13_July_2009&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/14_July_2009&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/15_July_2009&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/16_July_2009&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/17_July_2009&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/18_July_2009&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/19_July_2009&action=edit 19]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/20_July_2009&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/21_July_2009&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/22_July_2009&action=edit 22] [http://2009.igem.org/Team:Newcastle/Labwork/23_July_2009 23] [http://2009.igem.org/Team:Newcastle/Labwork/24_July_2009 24] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/25_July_2009&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/26_July_2009&action=edit 26]
[http://2009.igem.org/Team:Newcastle/Labwork/27_July_2009 27] [http://2009.igem.org/Team:Newcastle/Labwork/28_July_2009 28] [http://2009.igem.org/Team:Newcastle/Labwork/29_July_2009 29] [http://2009.igem.org/Team:Newcastle/Labwork/30_July_2009 30] [http://2009.igem.org/Team:Newcastle/Labwork/31_July_2009 31]
August
MTWTFSS
          [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/1_August_2009&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/2_August_2009&action=edit 2]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/3_August_2009&action=edit 3] [http://2009.igem.org/Team:Newcastle/Labwork/4_August_2009 4] [http://2009.igem.org/Team:Newcastle/Labwork/5_August_2009 5] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/6_August_2009&action=edit 6] [http://2009.igem.org/Team:Newcastle/Labwork/7_August_2009 7] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/8_August_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/9_August_2009&action=edit 9]
[http://2009.igem.org/Team:Newcastle/Labwork/10_August_2009 10] [http://2009.igem.org/Team:Newcastle/Labwork/11_August_2009 11] [http://2009.igem.org/Team:Newcastle/Labwork/12_August_2009 12] [http://2009.igem.org/Team:Newcastle/Labwork/13_August_2009 13] [http://2009.igem.org/Team:Newcastle/Labwork/14_August_2009 14] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/15_August_2009&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/16_August_2009&action=edit 16]
[http://2009.igem.org/Team:Newcastle/Labwork/17_August_2009 17] [http://2009.igem.org/Team:Newcastle/Labwork/18_August_2009 18] [http://2009.igem.org/Team:Newcastle/Labwork/19_August_2009 19] [http://2009.igem.org/Team:Newcastle/Labwork/20_August_2009 20] [http://2009.igem.org/Team:Newcastle/Labwork/21_August_2009 21] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/22_August_2009&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/23_August_2009&action=edit 23]
[http://2009.igem.org/Team:Newcastle/Labwork/24_August_2009 24] [http://2009.igem.org/Team:Newcastle/Labwork/25_August_2009 25] [http://2009.igem.org/Team:Newcastle/Labwork/26_August_2009 26] [http://2009.igem.org/Team:Newcastle/Labwork/27_August_2009 27] [http://2009.igem.org/Team:Newcastle/Labwork/28_August_2009 28] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/29_August_2009&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/30_August_2009&action=edit 30]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/31_August_2009&action=edit 31]
September
MTWTFSS
  [http://2009.igem.org/Team:Newcastle/Labwork/1_September_2009 1] [http://2009.igem.org/Team:Newcastle/Labwork/2_September_2009 2] [http://2009.igem.org/Team:Newcastle/Labwork/3_September_2009 3] [http://2009.igem.org/Team:Newcastle/Labwork/4_September_2009 4] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/5_September_2009&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/6_September_2009&action=edit 6]
[http://2009.igem.org/Team:Newcastle/Labwork/7_September_2009 7] [http://2009.igem.org/Team:Newcastle/Labwork/8_September_2009 8] [http://2009.igem.org/Team:Newcastle/Labwork/9_September_2009 9] [http://2009.igem.org/Team:Newcastle/Labwork/10_September_2009 10] [http://2009.igem.org/Team:Newcastle/Labwork/11_September_2009 11] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/12_September_2009&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/13_September_2009&action=edit 13]
[http://2009.igem.org/Team:Newcastle/Labwork/14_September_2009 14] [http://2009.igem.org/Team:Newcastle/Labwork/15_September_2009 15] [http://2009.igem.org/Team:Newcastle/Labwork/16_September_2009 16] [http://2009.igem.org/Team:Newcastle/Labwork/17_September_2009 17] [http://2009.igem.org/Team:Newcastle/Labwork/18_September_2009 18] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/19_September_2009&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/20_September_2009&action=edit 20]
[http://2009.igem.org/Team:Newcastle/Labwork/21_September_2009 21] [http://2009.igem.org/Team:Newcastle/Labwork/22_September_2009 22] [http://2009.igem.org/Team:Newcastle/Labwork/23_September_2009 23] [http://2009.igem.org/Team:Newcastle/Labwork/24_September_2009 24] [http://2009.igem.org/Team:Newcastle/Labwork/25_September_2009 25] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/26_September_2009&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/27_September_2009&action=edit 27]
[http://2009.igem.org/Team:Newcastle/Labwork/28_September_2009 28] [http://2009.igem.org/Team:Newcastle/Labwork/29_September_2009 29] [http://2009.igem.org/Team:Newcastle/Labwork/30_September_2009 30]
October
MTWTFSS
      [http://2009.igem.org/Team:Newcastle/Labwork/1_October_2009 1] [http://2009.igem.org/Team:Newcastle/Labwork/2_October_2009 2] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/3_October_2009&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/4_October_2009&action=edit 4]
[http://2009.igem.org/Team:Newcastle/Labwork/5_October_2009 5] [http://2009.igem.org/Team:Newcastle/Labwork/6_October_2009 6] [http://2009.igem.org/Team:Newcastle/Labwork/7_October_2009 7] [http://2009.igem.org/Team:Newcastle/Labwork/8_October_2009 8] [http://2009.igem.org/Team:Newcastle/Labwork/9_October_2009 9] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/10_October_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/11_October_2009&action=edit 11]
[http://2009.igem.org/Team:Newcastle/Labwork/12_October_2009 12] [http://2009.igem.org/Team:Newcastle/Labwork/13_October_2009 13] [http://2009.igem.org/Team:Newcastle/Labwork/14_October_2009 14] [http://2009.igem.org/Team:Newcastle/Labwork/15_October_2009 15] [http://2009.igem.org/Team:Newcastle/Labwork/16_October_2009 16] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/17_October_2009&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/18_October_2009&action=edit 18]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/19_October_2009&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/20_October_2009&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/21_October_2009&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/22_October_2009&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/23_October_2009&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/24_October_2009&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/25_October_2009&action=edit 25]
[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/26_October_2009&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/27_October_2009&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/28_October_2009&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/29_October_2009&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/30_October_2009&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/Labwork/31_October_2009&action=edit 31]



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