From 2008.igem.org
(Difference between revisions)
|
|
| Line 41: |
Line 41: |
| | |'''Miniprep''' | | |'''Miniprep''' |
| | |'''Biobrick''' | | |'''Biobrick''' |
| - | |'''Concentration (µg/mL)''' | + | |'''C° (µg/mL)''' |
| | |'''ratio 260/280''' | | |'''ratio 260/280''' |
| | |- | | |- |
Revision as of 16:03, 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
to be modified
Protocol Digestion
| Plasmid
| Description
| Miniprep used
| Enzymes
|
| MP3.3 ?
| B0015 (double terminator B0010-B0012) - FV
| 4
| EcoRI and XbaI
|
| MP119
| promoter J23101 - BV
| 1
| SpeI and PstI
|
| MP104
| PTet (Tet promoter) - BV
| 1
| SpeI and PstI
|
| MP114
| TetR - FI
| 1
| EcoRI and SpeI
|
| MP143
| gfp generator - BI
| 2
| SpeI and PstI
|
|
"
