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| - | [[Team:Paris/Notebook/Protocols#Digestion |protocol]] | + | [[Team:Paris/Notebook/Protocols#Digestion |Protocol]] |
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Revision as of 15:39, 19 August 2008
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Analysis of the cloning of OmpR and EnvZ
PCR screening
- positive control: S158 (pSB3K3)
- negative control: no template
- screening PCR programm
- elongation time: 1 min 30
- primers: O18 & O19
Reaction mixture (25 µL total volume)
- 12,5 µL Quick load PCR Mix
- 0,5 µL O18
- 0,5 µL O19
- 12 µL water
We wanted to analyse the transformants of the ligations L133 (OmpR in pSB1A2) and L134 (EnvZ in pSB1A2) from the experience of august 9th/11th.
3 clones of L133 and 3 clones of L134 were analysed.
Each clone tested was then cultured in 7,5 mL LB + ampicilline.
Miniprep and stock glycerol of L151
| Stock number
| Ligation name
| Biobricks
| Description
|
| S162.1
| L151.1
|   
| rbs-LasR - Double terminator
|
| S162.2
| L151.2
|
| S162.3
| L151.3
|
| Miniprep Name
| Ligation name
| Antibio
| Biobricks
| Description
|
| MP162.1
| L151.1
| Kan
|
| rbs-LasR - Double terminator
|
| MP162.2
| L151.2
|
| MP162.3
| L151.3
|
Digestion of MP162
Measurement of the concentration of MPs 162
| Miniprep Name
| Concentration (µg/mL)
| Ratio 260/280
|
| MP162.1
| 151
| 1.71
|
| MP162.2
| 172
| 1.73
|
| MP162.3
| 149
| 1.73
|
Digestion
Protocol
| Digestion name
| Template DNA
| Enzymes
| Volume of DNA
|
| D165
| MP162.2
| XbaI - PstI
| 5.81 µL
|
Ligation
Protocol
| Ligation Name
| Vector Name
| Volume Vector (µL)
| Insert
| Volume Insert (µL)
|
| L153
| D123.1
| 1.54
| D165
| 1.69
|
| L154
| D103.2
| 1.14
| D165
| 1.68
|
|
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