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:10/10/2017)... ... October 10, 2017 , ... The Pittcon Program Committee ... honoring scientists who have made outstanding contributions to analytical chemistry and applied ... the world’s leading conference and exposition for laboratory science, which will be held ...
(Date:10/9/2017)... ... October 09, 2017 , ... At its national board meeting in North ... in Harvard University’s Departments of Physics and Astronomy, has been selected for membership in ... winning team for the 2015 Breakthrough Prize in Fundamental physics for the discovery of ...
(Date:10/7/2017)... ... October 06, 2017 , ... ... instruments and applications consulting for microscopy and surface analysis, Nanoscience Instruments is ... Nanoscience Analytical offers a broad range of contract analysis services for advanced ...
(Date:10/6/2017)... , ... October 06, 2017 , ... ... within the healthcare and technology sector at their fourth annual Conference where founders, ... 30 inspiring speakers and the ELEVATE pitch competition showcasing early stage digital health ...
Breaking Biology Technology:
(Date:8/15/2017)... Aug. 15 2017   ivWatch LLC , a medical device ... therapy, today announced receipt of its ISO 13485 Certification, the global ... International Organization for Standardization (ISO®). ... ivWatch Model 400 Continuous Monitoring device for the early detection of ... "This is an important milestone ...
(Date:6/23/2017)... ITHACA, N.Y. , June 23, 2017 ... a leader in dairy research, today announced a new ... help reduce the chances that the global milk supply ... this dairy project, Cornell University has become the newest ... Food Supply Chain, a food safety initiative that includes ...
(Date:5/16/2017)... May 16, 2017  Veratad Technologies, LLC ( www.veratad.com ... age and identity verification solutions, announced today they will ... 2017, May 15 thru May 17, 2017, in ... International Trade Center. Identity impacts the ... in today,s quickly evolving digital world, defining identity is ...
Breaking Biology News(10 mins):