Acinetobacter ADP1

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(New page: == 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...)
(Transformations)
Line 6: Line 6:
== Transformations ==
== Transformations ==
 +
 +
* 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 ==
== Constructs ==

Revision as of 14:20, 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

  • 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
  • 2.2 in plasmid IMBB
  • 2.1-P1010-2.2 in plasmid pSB1AC3
  • 2.1-J04450-P1010-2.2 in plasmid pSB1AC3
Personal tools