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| - | {{Imperial/StartPage}}__NOTOC__ | + | {{Imperial/StartPage2}}__NOTOC__ |
| | {| cellpadding="10" border="0" | | {| cellpadding="10" border="0" |
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| | |{{#calendar: title=Imperial_College |year=2008 | month=08}} | | |{{#calendar: title=Imperial_College |year=2008 | month=08}} |
| | |{{#calendar: title=Imperial_College |year=2008 | month=09}} | | |{{#calendar: title=Imperial_College |year=2008 | month=09}} |
| - | | rowspan="2" bgcolor=#fff width="100%" | | + | | rowspan="2" bgcolor=#ffffff width="100%" | |
| | |} | | |} |
| | | | |
| - | =14th August 2008= | + | =14 August 2008= |
| | | | |
| | ==Wet Lab== | | ==Wet Lab== |
| | + | |
| | + | === Cloning === |
| | | | |
| | *Parts taken from the registry and transformed by electroporation into XL1-Blue ''E.coli'' along with XL1-Blue controls and grown on Kanamycin and Ampicillin plates. Parts taken: | | *Parts taken from the registry and transformed by electroporation into XL1-Blue ''E.coli'' along with XL1-Blue controls and grown on Kanamycin and Ampicillin plates. Parts taken: |
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| | **J04630 (GFP and Double Terminator) | | **J04630 (GFP and Double Terminator) |
| | **I13401 (mRFP and Double Terminator) | | **I13401 (mRFP and Double Terminator) |
| | + | |
| | + | ===''B.subtilis''=== |
| | + | *The transformation protocol 2 was carried out today, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol] |
| | + | *The basic principle of this protocol is that competent cells are prepared by growing a culture to a high O.D.<sub>600</sub> and then performing electroporation on these competent cells. |
| | + | *Today we grew the competent cells, we used an O.D.<sub>600</sub> of 1.5 before we harvested them. In addition electroporation was carried out using the plasmid pDR110. Previously we had mini-preped this to give a stock of 40ng/ul. We transformed with 40ng, 120ng, 200ng and 400ng in a volume of 10ul (water was used to make up to 10ul). |
| | + | *Transformed cells were plated out and placed into the 30<sup>o</sup>C incubator. |
| | | | |
| | ==Dry Lab== | | ==Dry Lab== |
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| | ==Microscope== | | ==Microscope== |
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| - | *Chris, Clinton, James and Prudence attended microscope training on Widefield 1, recording two 1 minute videos of ''B.subtilis'' swimming for analysis. | + | *Chris, Clinton, James and Prudence received microscope training on Widefield 1, recording two 60s videos of ''B.subtilis'' swimming for analysis |
| - | *Determined that best results would be obtained by bringing a ''B.subtilis'' overnight culture to the microscope facility and diluting 100 fold | + | *Determined that best results would be obtained by bringing a ''B.subtilis'' overnight culture to the microscope facility and diluting 100 fold. |
| | + | <br> |
| | + | {{Imperial/EndPage|Notebook|Notebook}} |
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| July
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| M | T | W | T | F | S | S
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| [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]
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| [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]
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| [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]
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| [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]
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| [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]
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| August
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| M | T | W | T | F | S | S
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| [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]
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| [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]
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| [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]
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| [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]
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| [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]
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| September
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| M | T | W | T | F | S | S
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| [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]
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| [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]
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| [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]
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| [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]
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| [http://2008.igem.org/Imperial_College/29_September_2008 29]
| [http://2008.igem.org/Imperial_College/30_September_2008 30]
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14 August 2008
Wet Lab
Cloning
- Parts taken from the registry and transformed by electroporation into XL1-Blue E.coli along with XL1-Blue controls and grown on Kanamycin and Ampicillin plates. Parts taken:
- C0012 (LacI)
- J31005 (Chloraphemicol acetyltransferase)
- B0015 (Double terminator)
- J04630 (GFP and Double Terminator)
- I13401 (mRFP and Double Terminator)
B.subtilis
- The transformation protocol 2 was carried out today, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol]
- The basic principle of this protocol is that competent cells are prepared by growing a culture to a high O.D.600 and then performing electroporation on these competent cells.
- Today we grew the competent cells, we used an O.D.600 of 1.5 before we harvested them. In addition electroporation was carried out using the plasmid pDR110. Previously we had mini-preped this to give a stock of 40ng/ul. We transformed with 40ng, 120ng, 200ng and 400ng in a volume of 10ul (water was used to make up to 10ul).
- Transformed cells were plated out and placed into the 30oC incubator.
Dry Lab
- Completed A More Complex Example of Bayesian Parameter Estimation of Tutorial 2.
- Sourced for better tracking algorithm, found SpotTracker which is more accurate than ParticleTracker.
- Went for microscope training, obtained 2 x videos on B.Subtilis motility, stored on server with James.
- Discussion with Dr. Suhail ( from Structural Bioinformatics Group,Imperial College London) on the computing requirements to analyse the cells motility. We will be allocated a linux 64-bit workstation, that we will be able to access remotely via SSH.
Microscope
- Chris, Clinton, James and Prudence received microscope training on Widefield 1, recording two 60s videos of B.subtilis swimming for analysis
- Determined that best results would be obtained by bringing a B.subtilis overnight culture to the microscope facility and diluting 100 fold.
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