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.



Page: All 1 2

Related biology technology :

1. Salmonella typhimurium
Post Your Comments:

(Date:11/30/2015)... 30, 2015 /PRNewswire/ - Zenith Epigenetics Corp. ("Zenith" or the ... C.W. Wong to its Board of Directors to replace ... with a wealth of experience as co-founder of Resverlogix, with ... --> --> Dr. Wong remarked, ... of directors. Zenith,s long standing expertise in epigenetics and the ...
(Date:11/30/2015)... -- TapImmune, Inc. (TPIV), a clinical-stage ... and gene-based immunotherapeutics and vaccines for the treatment of ... presenting at the 8 th Annual LD Micro ... PT. Dr. John N. Bonfiglio a TapImmune ... presentation and will join TapImmune management in meeting with ...
(Date:11/30/2015)... 2015  Northwest Biotherapeutics (NASDAQ: NWBO ) ("NW ... therapies for solid tumor cancers, announced today that the ... and the Company welcomes Neil Woodford,s call ... anonymous internet report on NW Bio.  The Company,s Board ... Powers stated, "We agree with Mr. Woodford that, ...
(Date:11/27/2015)... , ... November 27, 2015 , ... ... that includes over 2,000 technical presentations offered in symposia, oral sessions, workshops, ... applied spectroscopy, covers a wide range of applications such as, but not limited ...
Breaking Biology Technology:
(Date:11/4/2015)... York , November 4, 2015 ... a new market report published by Transparency Market Research "Home ... Growth, Trends and Forecast 2015 - 2022", the global home ... US$ 30.3 bn by 2022. The market is estimated ... forecast period from 2015 to 2022. Rising security needs ...
(Date:10/29/2015)... RESTON, Va. , Oct. 29, 2015 ... announced today that it has released a new version ... Daon customers in North America ... gains. IdentityX v4.0 also includes a FIDO UAF ... customers are already preparing to activate FIDO features. These ...
(Date:10/29/2015)... , Oct. 29, 2015 Today, ... a partnership with 2XU, a global leader in ... a smart hat with advanced bio-sensing technology. The ... athletes to monitor key biometrics to improve overall ... partnership, the two companies will bring together the most ...
Breaking Biology News(10 mins):