Acinetobacter ADP1: Difference between revisions
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== Constructs == | == Constructs == | ||
* 2.1 in plasmid IMBB | * 2.1 in plasmid IMBB | ||
** IMBB expresses TEM-1 β-lactamase and has a ColE1 origin | |||
* 2.2 in plasmid IMBB | * 2.2 in plasmid IMBB | ||
* 2.1-P1010-2.2 in plasmid pSB1AC3 | * 2.1-P1010-2.2 in plasmid pSB1AC3 | ||
* 2.1-J04450-P1010-2.2 in plasmid pSB1AC3 | * 2.1-J04450-P1010-2.2 in plasmid pSB1AC3 | ||
* gusA-specR reporter system | * gusA-specR reporter system |
Latest revision as of 12:28, 16 June 2009
Cultures
- ADP1 can be grown anywhere between room temperature and 37°C
- Will not survive at 4°C
- For long term storage, add 10% DMSO and store at -80°C
- Naturally resistant to ampicillin and chloramphenicol. Use spectinomycin, kanamycin, or tetracycline for selections.
Transformations
(derived from Metzgar et al. paper)
- Streak out cells on LB plate, grow overnight at 37°C
- Innoculate colonies into 2 mL LB and grow for 12-16 hours at 30°C with shaking
- Take a 20 μL aliquot and add to 280 μL fresh LB
- Add DNA
- Shake at 30°C for 2 hours
- Plate on appropriate media with antibiotic and let grow at 30°C
Constructs
- 2.1 in plasmid IMBB
- IMBB expresses TEM-1 β-lactamase and has a ColE1 origin
- 2.2 in plasmid IMBB
- 2.1-P1010-2.2 in plasmid pSB1AC3
- 2.1-J04450-P1010-2.2 in plasmid pSB1AC3
- gusA-specR reporter system