From 2008.igem.org
(Difference between revisions)
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Line 120: |
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| |D166 | | |D166 |
| |MP165.1 | | |MP165.1 |
- | |- FV | + | | - FV |
| |7.63 | | |7.63 |
| |17 | | |17 |
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Line 134: |
| |D131 | | |D131 |
| |MP122.2 | | |MP122.2 |
- | |- BV | + | | - BV |
| |10.2 | | |10.2 |
| |14.5 | | |14.5 |
| |XbaI and PstI | | |XbaI and PstI |
| |} | | |} |
Revision as of 16:17, 20 August 2008
← Yesterday ↓ Calendar ↑Tomorrow →
Screening of the cloning of E0240 and FlhDC+promotor
Spreading the clones in order to obtain single colonies
Strain
| Resistance
| Ligation
| DNA cloned
| vector
| expected size of the fragment amplified by VF & VR
| mesured size
|
S159.1
| kanamycine
| L139.1
| E0240 (GFP tripart)
| pSB3K3
| 1192 bp
| 1,5 kb 1,1 kb 0,6 kb
|
S161.1
| ampicilline
| L142.7
| FlhDC+promotor
| pSB1A2
| 1403 bp
| 1,4 0,4 kb 0,3 kb
|
The plates obtained from the speading of yesterday can't be used because there are not single colonies.
We have to try again, but with a stronger dilution of the bacteria or with a smaller volume of spreading.
Digestion
Aim : Construction of "pFlgA-RBS-YFP-dbl ter"
Measurement of concentration of minipreps
standard protocol
Miniprep
| Biobrick
| C° (µg/mL)
| ratio 260/280
|
MP164.1
| E0422
| 95
| 1.69
|
MP164.2
| E422
| 90
| 1.76
|
MP165.1
| E0430
| 131
| 1.74
|
MP165.2
| E0430
| nd
| nd
|
MP166.1
| E0432
| 112
| 1.65
|
MP166.2
| E0432
| 79
| 1.6
|
MP167.1
| E0420
| 199
| 1.72
|
MP167.2
| E0420
| 194
| 1.73
|
MP168.1
| I732078
| 111
| 1.65
|
MP168.2
| I732078
| 104
| 1.67
|
MP122.1
| E0840
| 56
| 1.58
|
MP122.2
| E0840
| 98
| 1.63
|
Digestion
Protocol Digestion
Name
| Miniprep used
| Description
| Vol MP (µl)
| Vol H2O (µl)
| Enzymes
|
D166
| MP165.1
| - FV
| 7.63
| 17
| EcoRI and XbaI
|
D167
| MP166.1
| - BV
| 8.9
| 15.8
| EcoRI and XbaI
|
D131
| MP122.2
| - BV
| 10.2
| 14.5
| XbaI and PstI
|
|