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- Protocol (see # 3) Experiments done by QIAcube
| Name
| Ligation
| Biobricks
| Description
|
| MP144
| L128.1
| pFlgA-RFP
D132 (FI) - D136 (FV)
|  
|
| L128.2
| pFlgA-RFP
D132 (FI) - D136 (FV)
|
|
| L128.3
| pFlgA-RFP
D132 (FI) - D136 (FV)
|
|
| L128.4
| pFlgA-RFP
D132 (FI) - D136 (FV)
|
|
| MP145
| L129.1
| pFlgB-RFP
D133 (FI) - D136 (FV)
|
|
| L129.2
| pFlgB-RFP
D133 (FI) - D136 (FV)
|
|
| L129.6
| pFlgB-RFP
D133 (FI) - D136 (FV)
|
|
| L129.7
| pFlgB-RFP
D133 (FI) - D136 (FV)
|
|
| MP146
| L130.1
| pFlhB-RFP
D134 (FI) - D136 (FV)
|
|
| L130.2
| pFlhB-RFP
D134 (FI) - D136 (FV)
|
|
| L130.7
| pFlhB-RFP
D134 (FI) - D136 (FV)
|
|
| L130.8
| pFlhB-RFP
D134 (FI) - D136 (FV)
|
|
Glycerol Stocks
| Strain
| Ligation
| Biobricks
| Description
|
| S143.1
| L128.1
| pFlgA-RFP
D132 (FI) - D136 (FV)
|
|
| S143.2
| L128.2
|
| S143.3
| L128.3
|
| S143.4
| L128.4
|
| S144.1
| L129.1
| pFlgB-RFP
D133 (FI) - D136 (FV)
|
|
| S144.2
| L129.2
|
| S144.6
| L129.6
|
| S144.7
| L129.7
|
| S145.1
| L130.1
| pFlhB-RFP
D134 (FI) - D136 (FV)
|
|
| S145.2
| L130.2
|
| S145.7
| L130.7
|
| S145.8
| L130.8
|
Results of the transformations we did yesterday
Concentration of the MiniPreps
| Miniprep
| Concentration (µg/mL)
| Ratio 260/280
|
| MP144
| 127
| 1.67
|
| MP145
| 166
| 1.66
|
| MP146
| 163
| 1.57
|
| MP147
| 154
| 1.63
|
| MP148
| 136
| 1.67
|
| MP149
| 151
| 1.63
|
| MP150
| 163
| 1.67
|
| MP151
| 188
| 1.57
|
| MP152
| 298
| 1.70
|
| MP153
| 142
| 1.64
|
| MP154
| 170
| 1.52
|
| MP155
| 143
| 1.55
|
New PCR screening with the right primers
Transformants with pFlgA, pFlgB and pFlhB cloned into J61002 are analysed by PCR but this time with the right primers: VF2 (O18) and VR (O19).
- template: bacteria from glycerol stock
- positive control: J61002 (with ptet-mRFP)
- negative control: no template
Electrophoresis
- 1% agarose gel
- 10 µL of each sample loaded
| well n°
| 1
| 2
| 3
| 4
| 5
| 6
| 7
| 8
| 9
| 10
| 11
| 12
| 13
| 14
| 15
| 16
|
| sample
| 1 kb DNA ladder
| positive control
| negative control
| L128 (pFlgA)
| L129 (pFlgB)
| L130 (pFlhB)
| 100 bp DNA ladder
|
| clone n°
|
| 1
| 2
| 3
| 4
| 1
| 2
| 6
| 7
| 1
| 2
| 7
| 8
|
|
| red fluorescence
|
| yes
| yes
| no
| no
| yes
| yes
| no
| no
| yes
| no
| no
| no
|
|
| expected size
|
| 1161 bp
| 0 bp
| 1339 bp
| 1339 bp
| 1338 bp
|
|
| measured size
|
| 1,2 kb
| 0 kb
| 1,2 kb
| 1,2 kb
| 1,3 kb
| 1,3 kb
| 1,2 kb
| 0 kb
| 1,4 kb
| 1,4 kb
| 1,2 kb
| 1,2 kb
| 1,2 kb
| 1,2 kb
|
|
|
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