|
|
| (6 intermediate revisions not shown) |
| Line 5: |
Line 5: |
| | |{{#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%" | |
| | |} | | |} |
| | = 20 August 2008 = | | = 20 August 2008 = |
| | | | |
| | == Wet Lab == | | == Wet Lab == |
| - | * Mini-prep | + | |
| | + | ===Cloning=== |
| | + | |
| | + | *GFP-3-mutB - Terminator and CAT overnight cultures miniprepped (all 3ml). |
| | + | *Single digests (''EcoRI'') of all registry sequences performed and assayed by gel to check vector size |
| | + | [[Image:20-8.PNG|center|frame| All run DNA was digested with ''EcoRI'' to linearise for optimal running. Lanes : M = Marker, 1 = Terminator Mini 1 , 2 = Terminator Mini 2, 3 = Terminator Mini 3, 4 = mRFP - terminator Mini 1 , 5 = mRFP - terminator Mini 2, 6 = mRFP - terminator Mini 3, 7 = AK3 Mini 1 , 8 = AK3 Mini 2, 9 = AK3 Mini 3, 10 = GFP-mut3b - terminator Mini 1 , 11 = GFP-mut3b - terminator Mini 2, 12 = GFP-mut3b - terminator Mini 3, 13 = CAT Mini 1 , 14 = CAT Mini 2, 15 = CAT Mini 3]] |
| | + | |
| | + | All major bands correspond to the approximate size of vector and insert. In several of the minis a second band is however visible is most likely to be denatured DNA as a byproduct of doing all the minipreps in one go (and so taking too long) |
| | | | |
| | ===''B.subtilis''=== | | ===''B.subtilis''=== |
| | *The transformation protocol 2, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol] was carried out on competent cells that had been grown to an O.D.<sub>600</sub> of 1.8. | | *The transformation protocol 2, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol] was carried out on competent cells that had been grown to an O.D.<sub>600</sub> of 1.8. |
| - | *The following concentrations of pDR110 DNA was used from the stock of 40ug/ul; 0, 40ng, 200ng, 400ng. | + | *The following concentrations of pDR110 DNA was used from the stock of 40ng/ul; 0, 40ng, 200ng, 400ng. One problem to note is that although the voltage and resistance can be controlled the pulse length cannot. The pulse length controls the length of time that the bacterium is exposed to the voltage. One problem is that if the pulse length drops below about 4msec, the efficiency of transformation is very low. In this set of transformations a range of 3msec to 10msec were seen. |
| | | | |
| | ==Dry Lab== | | ==Dry Lab== |
| | | | |
| - | * About to complete tutorial 2 | + | * About to complete Tutorial 2 |
| | * Preliminary work on validating the tracking algorithm written by EPFL using video generated from synthetic data | | * Preliminary work on validating the tracking algorithm written by EPFL using video generated from synthetic data |
| | * Preliminary work on growth curve of Bacillus Subtilis | | * Preliminary work on growth curve of Bacillus Subtilis |
| - | * We are aiming to produce a chassis characterisation | + | * We are aiming to produce a chassis characterisation. |
| | + | <br> |
| | + | {{Imperial/EndPage|Notebook|Notebook}} |
|
| July
|
| M | T | W | T | F | S | S
|
|
| [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
|
| 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]
|
|
|
20 August 2008
Wet Lab
Cloning
- GFP-3-mutB - Terminator and CAT overnight cultures miniprepped (all 3ml).
- Single digests (EcoRI) of all registry sequences performed and assayed by gel to check vector size
 All run DNA was digested with EcoRI to linearise for optimal running. Lanes : M = Marker, 1 = Terminator Mini 1 , 2 = Terminator Mini 2, 3 = Terminator Mini 3, 4 = mRFP - terminator Mini 1 , 5 = mRFP - terminator Mini 2, 6 = mRFP - terminator Mini 3, 7 = AK3 Mini 1 , 8 = AK3 Mini 2, 9 = AK3 Mini 3, 10 = GFP-mut3b - terminator Mini 1 , 11 = GFP-mut3b - terminator Mini 2, 12 = GFP-mut3b - terminator Mini 3, 13 = CAT Mini 1 , 14 = CAT Mini 2, 15 = CAT Mini 3 All major bands correspond to the approximate size of vector and insert. In several of the minis a second band is however visible is most likely to be denatured DNA as a byproduct of doing all the minipreps in one go (and so taking too long)
B.subtilis
- The transformation protocol 2, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol] was carried out on competent cells that had been grown to an O.D.600 of 1.8.
- The following concentrations of pDR110 DNA was used from the stock of 40ng/ul; 0, 40ng, 200ng, 400ng. One problem to note is that although the voltage and resistance can be controlled the pulse length cannot. The pulse length controls the length of time that the bacterium is exposed to the voltage. One problem is that if the pulse length drops below about 4msec, the efficiency of transformation is very low. In this set of transformations a range of 3msec to 10msec were seen.
Dry Lab
- About to complete Tutorial 2
- Preliminary work on validating the tracking algorithm written by EPFL using video generated from synthetic data
- Preliminary work on growth curve of Bacillus Subtilis
- We are aiming to produce a chassis characterisation.
|
"
