Paris/August 13
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< Paris
Revision as of 15:10, 16 August 2007 by David.bikard (Talk | contribs)
Digestions
- L1.2 (AraC-pBAD BV) Spe Pst
- L6.1 (rbs-CRE) Eco Pst
- L6.2 (rbs-CRE) Xba Pst
- L9 (lox66 FV) Eco Xba
- L10 (lox71 BV) Spe Pst
- MP8.1 (gfp tri-part FI) Eco Spe
digestion mix:
- 25µl plasmid
- 1µl BSA
- 10 µl NEB2
- 60µl H2O
- 2µl Enzyme1
- 2µl Enzyme2
Ligations
| 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 | |||
| C2 | D35 (L10 lox71 dig.SP) | 2µl | control of D35 digestion | |||
| C4 | D? (L9 lox66 EX) | 2µl | control of D? digestion | |||
| L26 | D33.2 (L6.2 RBS-Cre XP) | 5 | D? (L1 J61002::pbad SP) | 1 | Insertion of RBS-Cre behind araC/pBad | |
| L27 | D27 (ftsZ PCR XP) | 5 | D? (L1 J61002::pbad SP) | 1 | Insertion of ftsZ behind araC/pBad | |
| L28 | D32 (ftsAZ PCR XP) | 5 | D? (L1 J61002::pbad SP) | 1 | Insertion of ftsAZ behind araC/pBad | |
| L29 | D? (MP8.1 gfp-tripart ES) | 5 | D? (L9 pSB1A2::lox66 XP) | 2 | Insertion of gfp-tripart in front of lox66 | |
| L30(.1 & .2) | dapAp [O31+O32]0.2µM | 2 | D9.1 & D9.2 (dapAp [O31+O32]0.2µM) | 1 | cloning dapAp promotor in J61002 | |
Ligation mix:
- 2µl 10x buffer
- 1µl Ligase
- XµL plasmid DNA solution
- YµL insert DNA solution
- H2O qsp 20µL
Reaction in 20µl
ON @ 4°C
