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:6/23/2016)... ... June 23, 2016 , ... Charm Sciences, Inc. ... test has received AOAC Research Institute approval 061601. , “This is another AOAC-RI ... stated Bob Salter, Vice President of Regulatory and Industrial Affairs. “The Peel Plate ...
(Date:6/23/2016)... ON (PRWEB) , ... June 23, 2016 , ... STACS ... DNA Technical Leader at the Arkansas State Crime Laboratory, has joined STACS DNA as ... the STACS DNA team,” said Jocelyn Tremblay, President and COO of STACS DNA. “In ...
(Date:6/23/2016)... 2016 On Wednesday, June 22, 2016, ... 0.22%; the Dow Jones Industrial Average edged 0.27% lower to ... down 0.17%. Stock-Callers.com has initiated coverage on the following equities: ... (NASDAQ: NKTR ), Aralez Pharmaceuticals Inc. (NASDAQ: ... BIND ). Learn more about these stocks by accessing their ...
(Date:6/23/2016)... ReportsnReports.com adds 2016 global ... pharmaceuticals section with historic and forecast data along ... Complete report on the Cell Culture ... companies and supported with 261 tables and figures ... The Global Cell Culture Media Industry ...
Breaking Biology Technology:
(Date:6/2/2016)... --  The Weather Company , an IBM Business (NYSE: ... in which consumers will be able to interact with IBM ... voice or text and receive relevant information about the product ... have long sought an advertising solution that can create a ... and valuable; and can scale across millions of interactions and ...
(Date:5/20/2016)... 2016  VoiceIt is excited to announce its ... By working together, VoiceIt and VoicePass will ... VoicePass take slightly different approaches to voice biometrics, ... and usability. ... partnership. "This marketing and technology partnership ...
(Date:5/3/2016)... VILNIUS, Lithuania , May 3, 2016 /PRNewswire/ ... today released the MegaMatcher Automated Biometric Identification ... deployment of large-scale multi-biometric projects. MegaMatcher ABIS can ... and accuracy using any combination of fingerprint, face ... of MegaMatcher SDK and MegaMatcher ...
Breaking Biology News(10 mins):