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.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Brucella abortus
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/9/2016)... ... February 09, 2016 , ... Tunnell Consulting, Inc. announced that Frédéric Lefebvre ... will focus on acquiring new accounts and work closely with existing Tunnell clients throughout ... brings to our European clients more than 15 years of experience in the pharmaceutical ...
(Date:2/9/2016)... NJ (PRWEB) , ... February 09, 2016 , ... ... Dorman, former Vice President for Public Policy for the National Organization for Rare ... patient advocacy groups to ensure their voices are heard throughout the drug regulatory ...
(Date:2/9/2016)... and LONDON , February 9, 2016 /PRNewswire/ ... tech replace paper and protect IP   ... laboratory notebook (ELN) will be rolled out in ... and development (R&D) and protect valuable IP. Users will be ... a specific researcher or experiment as part of the application, ...
(Date:2/9/2016)... 9, 2016 DelveInsight,s, ... report provides in depth insights on the ... the Protein-Tyrosine Phosphatase 1B (PTP1B) Inhibitors. The ... various stages of development including Discovery, Pre-clinical, ... and Preregistration. Report covers the product clinical ...
Breaking Biology Technology:
(Date:2/3/2016)... 4, 2016 --> --> ... M (105.0), up 1,187% compared with fourth quarter of 2014. ... 517.6 M (loss: 30.0). Earnings per share increased to SEK ... 537.4 M (neg: 74.7). , --> ... to SEK 2,900.5 M (233.6), up 1,142% compared with 2014. ...
(Date:2/2/2016)... , Feb. 2, 2016 This BCC ... bioinformatic market by reviewing the recent advances in ... that drive the field forward. Includes forecast through ... Identify the challenges and opportunities that exist in ... software solution developers, as well as IT and ...
(Date:2/1/2016)... February 1, 2016 Rising sales ... drive global touchfree intuitive gesture control market ... Rising sales of consumer electronics coupled with new technological ... size through 2020   ... with new technological advancements to drive global touchfree intuitive ...
Breaking Biology News(10 mins):