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| | =23 September 2008= | | =23 September 2008= |
| | + | ==Wet Lab== |
| | + | ===Single Colony PCR=== |
| | + | [[Image:PCR-23.PNG|center|500px]] |
| | + | *In order to verify the ligations of parts (from geneart and PCR products) into the biobrick vector AK3 we have carried out a series of single colony PCRs using the primers Psb. |
| | + | *The conditions used were as follows: |
| | + | **1 cycle - 95<sup>o</sup>C for 30 seconds |
| | + | **30 cycles - 95<sup>o</sup>C for 30 seconds, 60<sup>o</sup>C for 30 seconds,72<sup>o</sup>C for 30 seconds |
| | + | **1 cycle - 72<sup>o</sup>C for 2 minutes |
| | + | *The numbers of the ligations correspond to the following ligation reactions: |
| | + | **6 L (from geneart construct 6) = LipA-Elastin |
| | + | **EpsE (from geneart construct 3) = EpsE gene |
| | + | **AmyE5' (from PCR products) |
| | + | **AmyE3' (from PCR products) |
| | + | **8<sub>1</sub> (from geneart constructs 6, PCR using mini DNA) = pGsiB-gsiB |
| | + | **8<sub>2</sub> (from geneart constructs 6, PCR using mini DNA) = pGsiB-gsiB |
| | + | **Negative contains no DNA, |
| | + | *The results show positive results for EpsE, Amy5',AmyE3', 8 (pGsiB-gsiB) |
| | + | *The result from AmyE3' is less clear, it appears that the 1st of the three is the correct size but all will be checked by mini-preping and then digestion. |
| | + | *There was no contamination in the negative control. |
| | + | |
| | + | ===Miniprep Digestion=== |
| | + | |
| | + | *All minpreps from yesterday digested with ''Eco''RI and ''Pst''I |
| | + | |
| | + | [[Image:Digest23-9.png|center|500px]] |
| | + | |
| | + | *Each lane is a separate miniprep (2 minipreps of each transformation). |
| | + | *Band from the EpsE digests is approximately correctly, although this is also approxiamtely the size of GFP-Termiantor and RFP-Terminator. In particular, the pSB1AK3 vector containing RFP-T was used to provide vector for the ligation. |
| | + | *All other mini-preps do not appear to have inserts! The gel had been imaged earlier and had also shown no evidence of inserts. |
| | + | *It should also be noted that the vector in all the minipreps appears to be the wrong size! |
| | | | |
| | ==Dry Lab== | | ==Dry Lab== |
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| | [[Image:V15Cell1.JPG|thumb|500px|center|Video 15 Cell 1]] | | [[Image:V15Cell1.JPG|thumb|500px|center|Video 15 Cell 1]] |
| | [[Image:V15Cell5.jpg|thumb|500px|center|Video 15 Cell 5]] | | [[Image:V15Cell5.jpg|thumb|500px|center|Video 15 Cell 5]] |
| | + | <br> |
| | + | {{Imperial/EndPage|Notebook|Notebook}} |
|
| August
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| M | T | W | T | F | S | S
|
|
|
|
|
| [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
|
| M | T | W | T | F | S | S
|
| [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]
|
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| October
|
| M | T | W | T | F | S | S
|
|
|
| [http://2008.igem.org/Imperial_College/1_October_2008 1]
| [http://2008.igem.org/Imperial_College/2_October_2008 2]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/3_October_2008&action=edit 3]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/4_October_2008&action=edit 4]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/5_October_2008&action=edit 5]
|
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_October_2008&action=edit 6]
| [http://2008.igem.org/Imperial_College/7_October_2008 7]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/8_October_2008&action=edit 8]
| [http://2008.igem.org/Imperial_College/9_October_2008 9]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/10_October_2008&action=edit 10]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/11_October_2008&action=edit 11]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/12_October_2008&action=edit 12]
|
| [http://2008.igem.org/Imperial_College/13_October_2008 13]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/14_October_2008&action=edit 14]
| [http://2008.igem.org/Imperial_College/15_October_2008 15]
| [http://2008.igem.org/Imperial_College/16_October_2008 16]
| [http://2008.igem.org/Imperial_College/17_October_2008 17]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/18_October_2008&action=edit 18]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/19_October_2008&action=edit 19]
|
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/20_October_2008&action=edit 20]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/21_October_2008&action=edit 21]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/22_October_2008&action=edit 22]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/23_October_2008&action=edit 23]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/24_October_2008&action=edit 24]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/25_October_2008&action=edit 25]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/26_October_2008&action=edit 26]
|
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/27_October_2008&action=edit 27]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/28_October_2008&action=edit 28]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/29_October_2008&action=edit 29]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/30_October_2008&action=edit 30]
| [http://2008.igem.org/wiki/index.php?title=Imperial_College/31_October_2008&action=edit 31]
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23 September 2008
Wet Lab
Single Colony PCR
- In order to verify the ligations of parts (from geneart and PCR products) into the biobrick vector AK3 we have carried out a series of single colony PCRs using the primers Psb.
- The conditions used were as follows:
- 1 cycle - 95oC for 30 seconds
- 30 cycles - 95oC for 30 seconds, 60oC for 30 seconds,72oC for 30 seconds
- 1 cycle - 72oC for 2 minutes
- The numbers of the ligations correspond to the following ligation reactions:
- 6 L (from geneart construct 6) = LipA-Elastin
- EpsE (from geneart construct 3) = EpsE gene
- AmyE5' (from PCR products)
- AmyE3' (from PCR products)
- 81 (from geneart constructs 6, PCR using mini DNA) = pGsiB-gsiB
- 82 (from geneart constructs 6, PCR using mini DNA) = pGsiB-gsiB
- Negative contains no DNA,
- The results show positive results for EpsE, Amy5',AmyE3', 8 (pGsiB-gsiB)
- The result from AmyE3' is less clear, it appears that the 1st of the three is the correct size but all will be checked by mini-preping and then digestion.
- There was no contamination in the negative control.
Miniprep Digestion
- All minpreps from yesterday digested with EcoRI and PstI
- Each lane is a separate miniprep (2 minipreps of each transformation).
- Band from the EpsE digests is approximately correctly, although this is also approxiamtely the size of GFP-Termiantor and RFP-Terminator. In particular, the pSB1AK3 vector containing RFP-T was used to provide vector for the ligation.
- All other mini-preps do not appear to have inserts! The gel had been imaged earlier and had also shown no evidence of inserts.
- It should also be noted that the vector in all the minipreps appears to be the wrong size!
Dry Lab
- Modelling Motility
- Done up the code to model the cell's trajectory using least square curve fit method.
- Fitted cell trajectories of 100908 Video 15. Results look promising:
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