Navigation Links
Salmonella typhimurium

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.532 01/2002 Microorganism Salmonella typhimurium LT2 Cell type Bacteria, gram negative Molecules injected Plasmid DNA (pBR322 in TE buffer) Growth medium LB medium Washing solution 10 mM HEPES, pH 7.0; 10% glycerol Electroporation solution 10% glycerol Outgrowth medium SOC medium (without antibiotics) Cuvette 2 mm gap width Reference Binotto J., et al 1991 Canadian Journal of Microbiology 37 474-477 Making electrocompetent cells:

1. Incoculate a flask of LB 1:100 with a fresh overnight culture, grow at 37 C with shaking to an O.D.640 of 0.75. Chill the cells in an ice-water bath for 15 min. 2. Harvest by centrifugation (4,000 x g, 10 min., 4 C). 3. Wash one time in the original culture volume with HEPES and one time with 10% glycerol using 1/100 of the original volume. 4. Resuspend in 10% glycerol.

Electroporation of cells:

  1. Add 2 l plasmid DNA (20 pg to 20 ng) to 40 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into a prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,400 V Time constant (T) 5 ms
  4. Immediately add 1 ml SOC medium and transfer to a sterile culture tube. Incubate for 1 hour at 37 C with shaking.
  5. Dilute the cells in SOC medium and plate on selective LB plates.
Expected Results: Transformation efficiency up to 4 x 108 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Salmonella typhimurium
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:12/6/2016)... , ... December 06, 2016 ... ... the Almac Group, the world’s largest privately-held contract pharmaceutical development and manufacturing ... Health, a leading biopharma outsourcing company combining a leading CRO and the ...
(Date:12/6/2016)... ... December 06, 2016 , ... Symbios Technologies, Inc. , ... company has engaged in a collaborative research partnership with Colorado State University (CSU) ... of the Vice President for Research. This agreement is designed to further the ...
(Date:12/6/2016)... -- Zimmer Biomet Holdings, Inc. (NYSE and SIX: ... of its previously-announced cash tender offers (the "Offers") ... accrued and unpaid interest to, but not including, ... expenses related to the Offers) (the "Maximum Tender ... table below (collectively, the "Notes"). The terms and ...
(Date:12/6/2016)... ... December 06, 2016 , ... ... management solutions headquartered in Aurora, Ohio, announced the opening of their new office ... the newly constructed facility is home to 200 employees focused on providing sales, ...
Breaking Biology Technology:
(Date:12/7/2016)... Avanade is helping Williams Martini Racing, one of ... biometric data in order to critically analyse every aspect ... against their rivals after their impressive, record-breaking pit stop ... with Williams during the 2016 season to capture and ... rate, temperature and peak acceleration) for key members of ...
(Date:12/6/2016)... Securus Technologies, a leading provider of ... safety, investigation, corrections and monitoring, and the Prison ... (5) year funding commitment by Securus to PEP ... and reentry support to more inmates and their ... the Prison Entrepreneurship Program (PEP) is an independent ...
(Date:11/30/2016)... Nov. 30, 2016 Not many of us realize that we spend ? ... so we need to do it well. Inadequate sleep levels have been found to ... stroke, diabetes, and even cancer. Maybe now is the best time to ... help them to manage their sleep quality? Continue Reading ... ...
Breaking Biology News(10 mins):