Paris/August 17
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Miniprep rxn
The following minipreps were performed:
- L26.1 & L26.2 (araC/pbad>>RBS-Cre)
- L27.2, L27.3, L27.4 & L27.5 (araC/pbad>>ftsZ)
- L28.2, L28.3 & L28.8 (araC/pBad>>ftsA-ftsZ)
- L29.1 & L29.2 (lox66-gfptripart)
- L30A1 & L30B1 (dapAp>>mRFP in J61002)
- MP1.1 (pJ23100) & MP2.1 (pJ23107)
Ligations
The following ligation reactions have been performed:
| Ligations | ||||||
|---|---|---|---|---|---|---|
| Number | Insert | Insert Volume (µL) | Vector | Vector Volume (µL) | Comments | Number of colonies |
| C1 | MP3.1 digested with EcoRI (D0) | 2 | control of ligation efficiency | |||
| C3 | D34 (lox66 BV) | 4µl | control of D34 digestion | |||
| C5 | D50 (BBa_J61002 BV) | 1.5µl | control of D50 digestion | |||
| C6 | D41 (araC/pBad-J61002 plasmid SP dig.) | 1µl | control of D41 digestion | |||
| C7 | D22 (pSB1A2 open vector EP) | 4µl | control of D22 digestion | |||
| C8 | D47 (pSB1A2 XP dig.) | 4µl | control of D47 digestion | |||
| C9 | D48 (pSB1A2 ES dig.) | 2µl | control of D48 digestion | |||
| L31 | D44 (B0015 terminator BI) | 4 | D35 (lox71 BV) | 3 | Insertion of B0015 terminator behind lox71 | |
| L32 | D45 (B0015 terminator FI) | 4 | D42 (Lox66 FV) | 3 | Insertion of B0015 terminator in front of lox66 | |
| L33 | D33.1 (RBS(B0030)CreORF BI) | 5 | D50 (BBa_J61002 BV) | 1.5 | pTet>>RBS-Cre | |
| L34 | D49 (lox71-gfptripart BI) | 5 | D50 (BBa_J61002 BV) | 1.5 | pTet>>lox71-gfptripart | |
| L35 | D49 (lox71-gfptripart BI) | 5 | D41 (araC/pBad-J61002 plasmid SP dig.) | 1 | araC/pBad>>lox71-gfptripart | |
| L36 | D13.4 (mRFP dig. SP) | 6 | D34 (lox66 BV) | 4 | lox66-mRFP | |
| L37 | D52 (RBS-dapAcoli BI) | 6 | D34 (lox66 BV) | 4 | lox66-RBS-dapAcoli BI | |
| L38 | D54 (RBS-dapAsubtilis BI) | 13 | D34 (lox66 BV) | 4 | lox66-RBS-dapAsubtilis BI | |
| L39 | D51 (RBS-dapAcoli BBinsert) | 4 | D22 (pSB1A2 open vector EP) | 4µl | RBS-dapAcoli cloned in pSB1A2 | |
| L40 | D53 (RBS-dapAsubtilis BBinsert) | 13 ( | D22 (pSB1A2 open vector EP) | 4µl | RBS-dapAsubtilis cloned in pSB1A2 | |
| L41 | D27' (lox71-ftsZ bw-insert) | 12 | D47 (pSB1A2 XP dig.) | 4µl | lox71-ftsZ cloned in pSB1A2 | |
| L42 | D32 (lox71-ftsAftsZ bw-insert) | 6 | D47 (pSB1A2 XP dig.) | 4µl | lox71-ftsA-Z cloned in pSB1A2 | |
| L43 | D55 (RBS-dapAsubtilis FI) | 15 | D48 (pSB1A2 ES dig.) | 2µl | RBS-dapAsubtilis cloned in pSB1A2 | |
Ligation mix:
- 2µl 10x buffer
- 1µl Ligase
- XµL plasmid DNA solution
- YµL insert DNA solution
- H2O qsp 20µL
Reaction in 20µl
ON @ 4°C
