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:2/5/2016)... N.C. , Feb. 5, 2016  In the ... role for a host of launch activities including the ... of this launch activity is especially high in the ... Best Practices and the Role of Medical ... help companies focused on oncology therapies find better ways ...
(Date:2/4/2016)... - New FDA action date of July 22, ... July 22, 2016   --> - ...   - Lifitegrast has the potential to be the ... signs and symptoms of dry eye disease in adults ... product approved in the U.S. in the past decade indicated for the treatment of ...
(Date:2/4/2016)... 2016  CytoSorbents Corporation (NASDAQ: CTSO ), ... CytoSorb® blood filter to treat deadly inflammation in ... announced that CEO Dr. Phillip Chan , ... Group,s 2016 Disruptive Growth & Healthcare Conference, providing ... Conference Presentation Details: Where: Convene Conference ...
(Date:2/4/2016)... Sinovac Biotech Ltd. ("Sinovac" or the "Company") ... products in China , today announced ... received on February 4, 2016 a preliminary non-binding proposal ... of PKU V-Ming ( Shanghai ) Investment ... Development ( Shenzhen ) Fund Management Co., ...
Breaking Biology Technology:
(Date:2/2/2016)... 2016 Checkpoint Inhibitors for Cancer – ... Are you interested in the future of cancer ... inhibitors. Visiongain,s report gives those predictions to 2026 ... level. Avoid falling behind in data or ... revenues those emerging cancer therapies can achieve. There ...
(Date:2/1/2016)... -- Today, the first day of American Heart Month, the ... first of its kind workplace health solution that leverages ... first application of Watson to ... Welltok will create a new offering that combines AHA,s ... on Welltok,s health optimization platform. The effort is intended ...
(Date:1/25/2016)... 25, 2016  Glencoe Software, the world-leading supplier of ... industries, will provide the data management solution OMERO Plus ... Photo - ... Phenotypic analysis measures the characteristics and ... comparisons between states such as health and disease, the ...
Breaking Biology News(10 mins):