From 2008.igem.org
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Revision as of 16:32, 20 August 2008
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← 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
| Template DNA
| 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
|
Protocol
| Well
| Sample
| Expected size
| Measured size
|
| 1
| 1kb ladder
|
|
|
| 2
| MP165.1
|
|
|
| 3
| MP166.1
|
|
|
| 4
| MP122.2
|
|
|
| 5
| no sample
|
|
|
| 6
| D166
|
|
|
| 7
| no sample
|
|
|
| 8
| D167
|
|
|
| 9
| no sample
|
|
|
| 10
| D131
|
|
|
| 11
| no sample
|
|
|
| 12
| 100pb ladder
|
|
|
|
"
