Imperial College/24 August 2008

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=24th August 2008=
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=24 August 2008=
==Wet Lab==
==Wet Lab==
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*Trial single colony PCR method to validate correct insertion into Biobricks
*Trial single colony PCR method to validate correct insertion into Biobricks
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===''B.subtilis''===
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=====Tuesday=====
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*Prepare 1x10 ml LB in 100ml flask in the morning and inoculate with ''B.subtilis'' in the afternoon. This culture will be used for the microscope tomorrow.
 +
*Perform the ''B.subtilis'' transformation 1 , [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4 click here for the protocol], this time use 500ul of competent cells as opposed to 200ul as we have previously used.
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=====Wednesday=====
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*Microscopy,
 +
*Check transformants,
 +
*Perform Colony PCR using on the transformed ''B.subtilis'' using primers to test for correct integration into the amyE site.
 +
 +
=====Thursday=====
 +
 +
*Perform integration with linear DNA from the pDR110 vector by cutting with suitable enzymes. We will use the transformation protocol 1.
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*''B.subtilis'' growth curve against O.D.<sub>600</sub>
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=====Friday=====
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*If colonies have grown from thursday transformation then perform colony PCR on these colonies to check correct integration of linear DNA.
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<br>
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{{Imperial/EndPage|Notebook|Notebook}}

Latest revision as of 20:39, 28 October 2008

July
MTWTFSS
  [http://2008.igem.org/wiki/index.php?title=Imperial_College/1_July_2008&action=edit 1] [http://2008.igem.org/wiki/index.php?title=Imperial_College/2_July_2008&action=edit 2] [http://2008.igem.org/wiki/index.php?title=Imperial_College/3_July_2008&action=edit 3] [http://2008.igem.org/wiki/index.php?title=Imperial_College/4_July_2008&action=edit 4] [http://2008.igem.org/wiki/index.php?title=Imperial_College/5_July_2008&action=edit 5] [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_July_2008&action=edit 6]
[http://2008.igem.org/Imperial_College/7_July_2008 7] [http://2008.igem.org/Imperial_College/8_July_2008 8] [http://2008.igem.org/Imperial_College/9_July_2008 9] [http://2008.igem.org/wiki/index.php?title=Imperial_College/10_July_2008&action=edit 10] [http://2008.igem.org/Imperial_College/11_July_2008 11] [http://2008.igem.org/wiki/index.php?title=Imperial_College/12_July_2008&action=edit 12] [http://2008.igem.org/wiki/index.php?title=Imperial_College/13_July_2008&action=edit 13]
[http://2008.igem.org/wiki/index.php?title=Imperial_College/14_July_2008&action=edit 14] [http://2008.igem.org/Imperial_College/15_July_2008 15] [http://2008.igem.org/Imperial_College/16_July_2008 16] [http://2008.igem.org/wiki/index.php?title=Imperial_College/17_July_2008&action=edit 17] [http://2008.igem.org/Imperial_College/18_July_2008 18] [http://2008.igem.org/wiki/index.php?title=Imperial_College/19_July_2008&action=edit 19] [http://2008.igem.org/wiki/index.php?title=Imperial_College/20_July_2008&action=edit 20]
[http://2008.igem.org/Imperial_College/21_July_2008 21] [http://2008.igem.org/Imperial_College/22_July_2008 22] [http://2008.igem.org/Imperial_College/23_July_2008 23] [http://2008.igem.org/Imperial_College/24_July_2008 24] [http://2008.igem.org/Imperial_College/25_July_2008 25] [http://2008.igem.org/wiki/index.php?title=Imperial_College/26_July_2008&action=edit 26] [http://2008.igem.org/wiki/index.php?title=Imperial_College/27_July_2008&action=edit 27]
[http://2008.igem.org/Imperial_College/28_July_2008 28] [http://2008.igem.org/Imperial_College/29_July_2008 29] [http://2008.igem.org/Imperial_College/30_July_2008 30] [http://2008.igem.org/wiki/index.php?title=Imperial_College/31_July_2008&action=edit 31]
August
MTWTFSS
        [http://2008.igem.org/Imperial_College/1_August_2008 1] [http://2008.igem.org/Imperial_College/2_August_2008 2] [http://2008.igem.org/wiki/index.php?title=Imperial_College/3_August_2008&action=edit 3]
[http://2008.igem.org/Imperial_College/4_August_2008 4] [http://2008.igem.org/Imperial_College/5_August_2008 5] [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_August_2008&action=edit 6] [http://2008.igem.org/Imperial_College/7_August_2008 7] [http://2008.igem.org/Imperial_College/8_August_2008 8] [http://2008.igem.org/wiki/index.php?title=Imperial_College/9_August_2008&action=edit 9] [http://2008.igem.org/Imperial_College/10_August_2008 10]
[http://2008.igem.org/Imperial_College/11_August_2008 11] [http://2008.igem.org/Imperial_College/12_August_2008 12] [http://2008.igem.org/Imperial_College/13_August_2008 13] [http://2008.igem.org/Imperial_College/14_August_2008 14] [http://2008.igem.org/Imperial_College/15_August_2008 15] [http://2008.igem.org/Imperial_College/16_August_2008 16] [http://2008.igem.org/Imperial_College/17_August_2008 17]
[http://2008.igem.org/Imperial_College/18_August_2008 18] [http://2008.igem.org/Imperial_College/19_August_2008 19] [http://2008.igem.org/Imperial_College/20_August_2008 20] [http://2008.igem.org/Imperial_College/21_August_2008 21] [http://2008.igem.org/Imperial_College/22_August_2008 22] [http://2008.igem.org/wiki/index.php?title=Imperial_College/23_August_2008&action=edit 23] [http://2008.igem.org/Imperial_College/24_August_2008 24]
[http://2008.igem.org/Imperial_College/25_August_2008 25] [http://2008.igem.org/Imperial_College/26_August_2008 26] [http://2008.igem.org/Imperial_College/27_August_2008 27] [http://2008.igem.org/Imperial_College/28_August_2008 28] [http://2008.igem.org/Imperial_College/29_August_2008 29] [http://2008.igem.org/wiki/index.php?title=Imperial_College/30_August_2008&action=edit 30] [http://2008.igem.org/Imperial_College/31_August_2008 31]
September
MTWTFSS
[http://2008.igem.org/Imperial_College/1_September_2008 1] [http://2008.igem.org/Imperial_College/2_September_2008 2] [http://2008.igem.org/Imperial_College/3_September_2008 3] [http://2008.igem.org/Imperial_College/4_September_2008 4] [http://2008.igem.org/Imperial_College/5_September_2008 5] [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_September_2008&action=edit 6] [http://2008.igem.org/Imperial_College/7_September_2008 7]
[http://2008.igem.org/Imperial_College/8_September_2008 8] [http://2008.igem.org/Imperial_College/9_September_2008 9] [http://2008.igem.org/Imperial_College/10_September_2008 10] [http://2008.igem.org/Imperial_College/11_September_2008 11] [http://2008.igem.org/Imperial_College/12_September_2008 12] [http://2008.igem.org/wiki/index.php?title=Imperial_College/13_September_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Imperial_College/14_September_2008&action=edit 14]
[http://2008.igem.org/Imperial_College/15_September_2008 15] [http://2008.igem.org/Imperial_College/16_September_2008 16] [http://2008.igem.org/Imperial_College/17_September_2008 17] [http://2008.igem.org/Imperial_College/18_September_2008 18] [http://2008.igem.org/Imperial_College/19_September_2008 19] [http://2008.igem.org/wiki/index.php?title=Imperial_College/20_September_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=Imperial_College/21_September_2008&action=edit 21]
[http://2008.igem.org/Imperial_College/22_September_2008 22] [http://2008.igem.org/Imperial_College/23_September_2008 23] [http://2008.igem.org/Imperial_College/24_September_2008 24] [http://2008.igem.org/Imperial_College/25_September_2008 25] [http://2008.igem.org/Imperial_College/26_September_2008 26] [http://2008.igem.org/wiki/index.php?title=Imperial_College/27_September_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=Imperial_College/28_September_2008&action=edit 28]
[http://2008.igem.org/Imperial_College/29_September_2008 29] [http://2008.igem.org/Imperial_College/30_September_2008 30]

24 August 2008

Wet Lab

Cloning

Tuesday
  • Prepare plates growth characterisation later in week.
  • Prepare overnight cultures of the Terminator, Chlormaphemicol Acetyltransferase, mRFP - Terminator and GFP-3-mutB - Terminator in XL1-Blue for midiprepping on Wednesday.
  • Run trial PCR reactions for genomic template and/or pDR111 template PCR cloning steps and validate by fragment size on gel
Wednesday
  • Run PCR cloning reactions for those not done on Tuesday adn validate as before
Thursday
  • Cut PCR clones, ligate into Biobricks adn transform XL1-Blue E.coli
Friday
  • Trial single colony PCR method to validate correct insertion into Biobricks

B.subtilis

Tuesday
  • Prepare 1x10 ml LB in 100ml flask in the morning and inoculate with B.subtilis in the afternoon. This culture will be used for the microscope tomorrow.
  • Perform the B.subtilis transformation 1 , [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4 click here for the protocol], this time use 500ul of competent cells as opposed to 200ul as we have previously used.
Wednesday
  • Microscopy,
  • Check transformants,
  • Perform Colony PCR using on the transformed B.subtilis using primers to test for correct integration into the amyE site.
Thursday
  • Perform integration with linear DNA from the pDR110 vector by cutting with suitable enzymes. We will use the transformation protocol 1.
  • B.subtilis growth curve against O.D.600
Friday
  • If colonies have grown from thursday transformation then perform colony PCR on these colonies to check correct integration of linear DNA.



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