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:10/12/2017)... ... ... AMRI, a global contract research, development and manufacturing organization ... quality of life, will now be offering its impurity solutions as a stand-alone ... for all new drug products, including the finalization of ICH M7 earlier this ...
(Date:10/11/2017)... CA (PRWEB) , ... October 11, 2017 , ... ... upregulate any gene in its endogenous context, enabling overexpression experiments and avoiding the ... system with small RNA guides is transformative for performing systematic gain-of-function studies. ...
(Date:10/11/2017)... , ... October 11, 2017 , ... ... announced today it will be hosting a Webinar titled, “Pathology is going digital. ... Associates , on digital pathology adoption best practices and how Proscia improves lab ...
(Date:10/11/2017)... ... October 11, 2017 , ... Disappearing forests and increased emissions are the main ... people each year. Especially those living in larger cities are affected by air pollution ... of the most pollution-affected countries globally - decided to take action. , “I knew ...
Breaking Biology Technology:
(Date:3/30/2017)... 30, 2017 The research team of The ... (3D) fingerprint identification by adopting ground breaking 3D fingerprint minutiae recovery ... of speed and accuracy for use in identification, crime investigation, immigration ... ... A research team ...
(Date:3/28/2017)... 28, 2017 The report "Video ... Monitors, Servers, Storage Devices), Software (Video Analytics, VMS), and ... Global Forecast to 2022", published by MarketsandMarkets, the market ... is projected to reach USD 75.64 Billion by 2022, ... The base year considered for the study is 2016 ...
(Date:3/24/2017)... Research and Markets has announced the addition of the "Global ... to 2025" report to their offering. ... The Global Biometric Vehicle Access System Market ... the next decade to reach approximately $1,580 million by 2025. ... for all the given segments on global as well as regional ...
Breaking Biology News(10 mins):