From 2008.igem.org
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| | ==='''Protocol'''=== | | ==='''Protocol'''=== |
| | ==='''Results'''=== | | ==='''Results'''=== |
| - | ==='''Digestions'''===
| + | =='''Digestions'''== |
| | | | |
| | =='''Digestion'''== | | =='''Digestion'''== |
Revision as of 14:28, 16 August 2008
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← Yesterday ↓ Calendar ↑Tomorrow →
Transformation of the ligations we did yesterday
We transformed L 143, L 144 and the negative controls T1 and T2, using Invitrogen's TOP10 chemically competent cells standard protocol.
PCR amplification of flhDC and its promoter
List of PCRs
Protocol
Results
Digestions
Digestion
Measurement of concentration of minipreps
standard protocol
| Digestion
| Miniprep used
| Concentration (µg/mL)
| ratio 260/280
|
| D146
| MP148.2
| 123
| 1.58
|
| D147
| MP153.3
| 113
| 1.68
|
Digestion
Protocol Digestion
Ligation
Protocol Ligation
| Ligation name
| Insert
| Vector
|
| L150
| D146 (strongest rbs-TetR-GFP tripart)
| D105 (pLas)
|
| L151
| D147 (strongest rbs-LasR activator with LVA)
| D125 (Double terminator)
|
| Control 1
| /
| D105
|
| Control 2
| /
| D125
|
| Positive Control
| Puc19
|
Analysis of yesterday PCR screening
Starting the construction of the Promoter characterization plasmid
Digestion
Measurement of concentration of minipreps
standard protocol
| Plasmid
| Miniprep
| Concentration (µg/mL)
| ratio 260/280
|
| MP3
| 3
| 152
| 1.43
|
| MP3
| 4
| 775
| 1.21
|
| MP101
| 1
| 317
| 1.66
|
| MP101
| 2
| 389
| 1.36
|
| MP101
| 4
| 209
| 1.76
|
| MP104
| 1
| 173
| 1.32
|
| MP104
| 3
| 43
| 1.85
|
| MP104
| 4
| 52
| 1.66
|
| MP114
| 1
| 173
| 1.75
|
| MP114
| 2
| 263
| 1.43
|
| MP143
| 1
| 133
| 1.55
|
| MP143
| 2
| 132
| 1.70
|
Digestion
Protocol Digestion
| Plasmid
| Description
| Miniprep used
| Enzymes
|
| MP3
| B0015 (double terminator B0010-B0012)
| 4
| EcoRI and XbaI
|
| MP114
| TetR
| 1
| EcoRI and SpeI
|
| MP104
| PTet (Tet promoter)
| 1
| SpeI and PstI
|
| MP101
| promoter J23101
| 1
| SpeI and PstI
|
| MP143
| gfp generator
| 2
| SpeI and PstI
|
|
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