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:4/26/2017)... Maryland (PRWEB) , ... April 26, 2017 , ... ... will be demonstrating its new Bioflash MailGuardtm mail security screening solution at the ... The Bioflash MailGuard system provides a fast, highly accurate, easy to use ...
(Date:4/26/2017)... , April 26, 2017  Genisphere LLC, ... platform, has signed a collaborative and sponsored research ... Silvia Muro . The overall goal of ... pharmacodynamics of various 3DNA designs and formulations after ... targeting diseases of the vasculature as well as ...
(Date:4/25/2017)... Diego, CA (PRWEB) , ... April 25, 2017 ... ... division of L3 Healthcare, is pleased to announce the company is now a ... management. The iMedNet software certification enables the company’s clinical research team to ...
(Date:4/25/2017)... ... April 25, 2017 , ... Covalent Metrology ... Analytical Services unit provides high-quality data to clients, both faster and cheaper ... receipt. There are no price premiums, and customers are welcome to participate in ...
Breaking Biology Technology:
(Date:3/23/2017)... , Mar. 23, 2017 Research and ... System Market Analysis & Trends - Industry Forecast to 2025" ... ... grow at a CAGR of around 8.8% over the next decade ... industry report analyzes the market estimates and forecasts for all the ...
(Date:3/22/2017)... , March 21, 2017 Optimove ... used by retailers such as 1-800-Flowers and AdoreMe, ... — Product Recommendations and Replenishment. Using Optimove,s machine learning ... personalized product and replenishment recommendations to their customers ... on predictions of customer intent drawn from a ...
(Date:3/20/2017)... Germany , March 20, 2017 At this ... Hamburg -based biometrics manufacturer DERMALOG. The Chancellor came to the DERMALOG ... is this year,s CeBIT partner country. At the largest German ... biometrics in use: fingerprint, face and iris recognition as well as DERMALOG´s ... ...
Breaking Biology News(10 mins):