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:6/23/2016)... 23, 2016 Houston Methodist Willowbrook Hospital ... Sports Association to serve as their official health ... Methodist Willowbrook will provide sponsorship support, athletic training ... association coaches, volunteers, athletes and families. ... Sports Association and to bring Houston Methodist quality ...
(Date:6/23/2016)... ... June 23, 2016 , ... Supplyframe, the Industry Network for ... Design Lab . Located in Pasadena, Calif., the Design Lab’s mission is to ... are designed, built and brought to market. , The Design Lab is Supplyframe’s ...
(Date:6/23/2016)... Andrew D Zelenetz ... Published recently in Oncology ... touchONCOLOGY, Andrew D Zelenetz , discusses the ... is placing an increasing burden on healthcare systems ... With the patents on many biologics expiring, interest ...
(Date:6/23/2016)... -- Amgen (NASDAQ: AMGN ) today announced a ... sciences incubator to accelerate the development of new therapies ... QB3@953 was created to help high-potential life science and ... stage organizations - access to laboratory infrastructure. ... "Amgen Golden Ticket" awards, providing each winner with one ...
Breaking Biology Technology:
(Date:5/9/2016)... DUBAI , UAE, May 9, 2016 ... choice when it comes to expanding freedom for high ... Even in today,s globally connected world, there ... online conferencing system could ever duplicate sealing your deal ... are obtaining second passports by taking advantage of citizenship ...
(Date:4/28/2016)... and BANGALORE, India , April 28, ... Systems, a product subsidiary of Infosys (NYSE: INFY ... announced a global partnership that will provide end ... use mobile banking and payment services.      (Logo: ... key innovation area for financial services, but it also plays ...
(Date:4/26/2016)... and LONDON , April ... part of EdgeVerve Systems, a product subsidiary of ... today announced a partnership to integrate the Onegini ...      (Logo: http://photos.prnewswire.com/prnh/20151104/283829LOGO ) ... their customers enhanced security to access and transact ...
Breaking Biology News(10 mins):