From 2008.igem.org
(Difference between revisions)
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| |'''Ligation name''' | | |'''Ligation name''' |
| |'''Insert name ''' | | |'''Insert name ''' |
- | |'''Volume of insert µL''' | + | |'''V<sub>insert</sub> µL''' |
| |'''Vector name''' | | |'''Vector name''' |
- | |'''Volume of Vector µL''' | + | |'''V<sub>Vector</sub> µL''' |
| |- | | |- |
| |L 143 (Kan) | | |L 143 (Kan) |
Revision as of 14:52, 17 August 2008
← Yesterday ↓ Calendar ↑Tomorrow →
Analysis of the transformation we did yesterday
L143, L144, T1 and T2 showed no colonies. The positive control with pUC19 worked well.
We suppose that the ligation did not work. We will do it again today.
Ligation
Ligation
Cleaning of the DNA after the digestion
We used the QIAcube to wash the DNA, following the standard protocol.
Measure of DNA concentration of the digestion products
We used the biophotometer.
Settings:
- 10 µL of template DNA in 50 µL of pure water
- Blank : 10 µL of EB buffer in 50 µL of water.
Digestion name
| What's in ?
| DNA concentration (ng/µL)
|
D 149
| pfliL
| 2
|
D 150
| pfliL
| 3
|
D 151
| pSB3K3
| 12
|
D 152
| pSB3K3
| 20
|
D 153
| gene flhDC
| 7
|
D 154
| gene flhDC
| 12
|
D 155
| pflhDC
| 16
|
D 136
| j61002
| 12
|
D 145
| pSB1A2
| 21
|
List of ligations
- We ligated the DNA following the standard protocol.
- T1, T2, T3 and T4 are the autoligation controls for L 143, L144, L145 and L147
Ligation name
| Insert name
| Vinsert µL
| Vector name
| VVector µL
|
L 143 (Kan)
| D 149 (pfliL)
| 8
| D 137 (pSB3K3)
| 4
|
L 144 (Kan)
| D 150 (pfliL)
| 5
| D 152 (pSB3K3)
| 2.5
|
L 145 (Amp)
| D 153 (g flhDC)
| 10
| D 145 (pSB1A2)
| 2.5
|
L 146 (Amp)
| D 154 (g flhDC)
| 5
| D 145 (pSB1A2)
| 2.5
|
L 147 (Amp)
| D 155 (p flhDC)
| 1
| D 136 (J61002)
| 4
|
Protocol
Transformation
Transformation protocol
Name
| Description
| Antibio
|
Ligations
|
L150
| D105(BV) - D146(BI) pLas - strongest rbs-TetR-GFP tripart
| Amp
|
L151
| D147(FI) - D125 (FV) strongest rbs-LasR activator with LVA - Double terminator
| Kan
|
Controls
|
C1
| D105
| Amp
|
C2
| D125
| Kan
|
Positive Control
| pUC19
| Amp
|
|