Team:Newcastle/Labwork/14 August 2009

From 2009.igem.org

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=Lab 14/08/09=
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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 - 14th August 2009=
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[[Image:Team Newcastle 2009 iGEM 14-08-09 IMG 0394.JPG|350px|center]]
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=<font color="Orange"><u>Overview</u></font>=
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<font color="Orange">
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*[[#Metal Sensor Team|Metal Sensor Team]] '''- prepared some LB + Chloramphenicol + starch plates and added ''pGFP-rrnB'' ''B. subtilis'' transformants to it '''
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<br>
==<u>Metal Sensor Team</u>==
==<u>Metal Sensor Team</u>==
===Introduction and Summary===
===Introduction and Summary===
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So far we have shown that our ''Bacillus subtilis 168'' cultures, once the ''GFP-rrnb'' DNA has been added, can survive on LB + Chloramphenicol media. This suggests that we have successfully transformed our ''B. subtilis'' cells as the wild type ''B. subtilis'' cells do not have this resistance. However we need to prove that the resistance inherited by the ''Bacillus subtilis'' bacteria is due to the ''GFP-rrnb'' and not acquired by other means. '''Today, we intend to pour some LB + Chloramphenicol + starch plates and then plate our 'transformed' ''Bacillus'' cells onto these plates.''' The reason for adding starch to the plates: the ''GFP-rrnb'' plasmid removes the ''Bacillus subtilis'' bacteria's ability to break down starch using the ''amyE'' enzyme - starch plates will prove the bacteria have been transformed.
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So far we have shown that our ''Bacillus subtilis 168'' cultures, once the ''pGFP-rrnB'' DNA has been added, can survive on LB + Chloramphenicol media. This suggests that we have successfully transformed our ''B. subtilis'' cells as the wild type ''B. subtilis'' cells do not have this resistance. However we need to prove that the resistance inherited by the ''Bacillus subtilis'' bacteria is due to the ''GFP-rrnb'' and not acquired by other means. '''Today, we intend to pour some LB + Chloramphenicol + starch plates and then plate our 'transformed' ''Bacillus'' cells onto these plates.''' The reason for adding starch to the plates: the ''GFP-rrnb'' plasmid removes the ''Bacillus subtilis'' bacteria's ability to break down starch using the ''amyE'' enzyme - starch plates will prove the bacteria have been transformed.
===Changes to protocol===
===Changes to protocol===
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===Work we did===
===Work we did===
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[[Image:Team Newcastle iGEM 2009 14-08-09 IMG 0403.JPG|thumb|200px|Two 1 litre flasks - one containing 500ml LB + agar with starch also present and the other flask containing 500ml of deionised water]]
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[[Image:Team Newcastle iGEM 2009 14-08-09 IMG 0421.JPG|thumb|200px|Mathew pouring the LB starch + chloramphenicol plates under aseptic technique]]
* Using the [https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/Making_Agar_Plates#Making_up_starch_plates 'pouring plates' protocol], we made up 1 litre of LB + agar + starch solution and once it had been autoclaved and properly prepared (with chloramphenicol - this is the resistance displayed in ''GFP-rrnB''), the solution was poured into plates.
* Using the [https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/Making_Agar_Plates#Making_up_starch_plates 'pouring plates' protocol], we made up 1 litre of LB + agar + starch solution and once it had been autoclaved and properly prepared (with chloramphenicol - this is the resistance displayed in ''GFP-rrnB''), the solution was poured into plates.
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* The plate was then left in the 37ºC incubator overnight.
* The plate was then left in the 37ºC incubator overnight.
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== Chassis team ==
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{{:Team:Newcastle/Project/Labwork/CalTemplate}}
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[[Image:Newcastle 14 August BSub Transformation test 50.jpg|thumb|The test transformation 50ul plate.]]
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[[Image:Newcastle 14 August BSub Transformation test 200.jpg|thumb|The test transformation 200ul plate.]]
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[[Image:Newcastle 14 August BSub Transformation test control.jpg|thumb|The test transformation control plate.]]
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[[Image:Newcastle 14 August BSub Transformation test h2o.jpg|thumb|The test transformation water plate.]]
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[[Image:Newcastle 14 August cwlD plate 1.jpg|thumb|The cwlD plate.]]
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[[Image:Newcastle 14 August cwlD plate 101.jpg|thumb|The cwlD 10<sup>-1</sup> plate.]]
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[[Image:Newcastle 14 August cwlD plate 102.jpg|thumb|The cwlD 10<sup>-2</sup> plate.]]
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[[Image:Newcastle 14 August cwlD plate 103.jpg|thumb|The cwlD 10<sup>-3</sup> plate.]]
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[[Image:Newcastle 14 August cwlD plate 104.jpg|thumb|The cwlD 10<sup>-4</sup> plate.]]
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[[Image:Newcastle 14 August cwlD plate 105.jpg|thumb|The cwlD 10<sup>-5</sup> plate.]]
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[[Image:Newcastle 14 August cwlD plate notreat.jpg|thumb|The no treatment cwlD plate.]]
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{{:Team:Newcastle/Footer}}
{{:Team:Newcastle/Footer}}
{{:Team:Newcastle/Right}}
{{:Team:Newcastle/Right}}

Latest revision as of 12:06, 20 August 2010


Team Newcastle 2009 iGEM ProbationaryP-Sign.PNG

Formal Lab Session - 14th August 2009

Team Newcastle 2009 iGEM 14-08-09 IMG 0394.JPG

Overview

  • Metal Sensor Team - prepared some LB + Chloramphenicol + starch plates and added pGFP-rrnB B. subtilis transformants to it


Metal Sensor Team

Introduction and Summary

So far we have shown that our Bacillus subtilis 168 cultures, once the pGFP-rrnB DNA has been added, can survive on LB + Chloramphenicol media. This suggests that we have successfully transformed our B. subtilis cells as the wild type B. subtilis cells do not have this resistance. However we need to prove that the resistance inherited by the Bacillus subtilis bacteria is due to the GFP-rrnb and not acquired by other means. Today, we intend to pour some LB + Chloramphenicol + starch plates and then plate our 'transformed' Bacillus cells onto these plates. The reason for adding starch to the plates: the GFP-rrnb plasmid removes the Bacillus subtilis bacteria's ability to break down starch using the amyE enzyme - starch plates will prove the bacteria have been transformed.

Changes to protocol

There were no changes to the protocol described here except for the fact that the chosen antibiotic is chloramphenicol

Work we did

Two 1 litre flasks - one containing 500ml LB + agar with starch also present and the other flask containing 500ml of deionised water
Mathew pouring the LB starch + chloramphenicol plates under aseptic technique
  • Using the 'pouring plates' protocol, we made up 1 litre of LB + agar + starch solution and once it had been autoclaved and properly prepared (with chloramphenicol - this is the resistance displayed in GFP-rrnB), the solution was poured into plates.


  • These plates were then left on the bench for 1 hour. Three plates were taken from the stack and placed in the 42ºC incubator to dry off.


  • Once the plates had dried, a grid of 46 squares was drawn onto the base of the plate. These squares were then numbered from 1-46.


  • Within square 1, a mark was made in the agar surface (in the shape of an 'X')with wild type Bacillus subtilis.


  • Within squares 2-46, marks were also made on the agar surface but this time it was with our transformed Bacillus subtilis (which should have taken up gfp-rrnb)


  • The plate was then left in the 37ºC incubator overnight.
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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