Navigation Links
Brucella abortus

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.508 12/2001 Microorganism Brucella abortus Cell type Bacteria, gram negative Molecules injected Plasmid DNA (pBA.sodknr) Growth medium Trypticase soy broth Washing solution Sterile, cold water Electroporation solution 10% glycerol Outgrowth medium Trypticase soy broth Cuvette 2 mm gap width Reference Tatum F.M., et al 1992 Infection and Immunity 60 2863-2869

Making electrocompetent cells:

  1. Grow cells at 30 C to a density of O.D.600: 1.5-2.0.

  2. Harvest by centrifugation and wash five to seven times in sterile, cold water.

  3. Resuspend the cells in 10% glycerol to a final concentration of 4-6 x 1011 cell/ml and store frozen at 70 C.

  4. For electroporation frozen cells were thawed in ice and diluted with sterile water to a density of 1 x 1010 cells/ml prior to use.

Electroporation of cells:

  1. Grow cells in Trypticase soy broth at 37 C with vigorous shaking, chill on ice for 10 min.
  2. Harvest by centrifugation (10 min, 10,000 x g). Wash three times in an equal volume of sterile cold water.
  3. Resuspend cells in 1/500 volume of 10% glycerol (number of cells: 1010 cells/ml). Freeze aliquots on dry ice and
    store at 70 C.

    Mode Prokaryotes O Voltage (V) 2,500 V Time constant (T) 5 ms
  4. Immediately add 1 ml trypticase soy broth, incubate 6 hours at 37 C.
  5. Plate cells on selective tryptose agar plates, incubate at 37 C for four days.



Page: All 1 2

Related biology technology :

1. Brucella abortus
Post Your Comments:

(Date:11/24/2015)... , Nov. 24, 2015 /PRNewswire/ - Aeterna ... announced today that the remaining 11,000 post-share consolidation ... Purchase Warrants (the "Series B Warrants") subject to ... exercised on November 23, 2015, which will result ... After giving effect to the issuance of such ...
(Date:11/24/2015)... ... November 24, 2015 , ... ... are paramount. Insertion points for in-line sensors can represent a weak spot where ... InTrac 781/784 series of retractable sensor housings , which are designed to tolerate ...
(Date:11/24/2015)... Vancouver, BC (PRWEB) , ... November 24, 2015 ... ... to our customer, OrthoAccel® Technologies, Inc., on being named to Deloitte's 2015 Technology ... Creation Technologies’ Texas facility, OrthoAccel manufactures AcceleDent®, a FDA-cleared, Class II medical device ...
(Date:11/24/2015)... 24, 2015 Capricor Therapeutics, Inc. ... the discovery, development and commercialization of first-in-class therapeutics, today ... Officer, is scheduled to present at the 2015 Piper ... a.m. EST, at The Lotte New York Palace Hotel ... . --> . ...
Breaking Biology Technology:
(Date:11/12/2015)... -- A golden retriever that stayed healthy despite having the ... a new lead for treating this muscle-wasting disorder, report ... MIT and Harvard and the University of São Paolo ... Cell, pinpoints a protective gene that boosts muscle ... Boston Children,s lab of Lou Kunkel , PhD, ...
(Date:11/10/2015)... , Nov. 10, 2015 ... behavioral biometrics that helps to identify and verify ... Signature is considered as the secure and accurate ... identification of a particular individual because each individual,s ... accurate results especially when dynamic signature of an ...
(Date:10/29/2015)... 2015   MedNet Solutions , an innovative SaaS-based ... clinical research, is pleased to announce that it has ... as one of only three finalists for a ... and Growing" category. The Tekne Awards honor Minnesota ... technology innovation and leadership. iMedNet™ eClinical ...
Breaking Biology News(10 mins):