Navigation Links
Streptococcus salivarius

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.541 03/2002 Microorganism Streptococcus salivarius Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium HJGS (Hogg-Jago glucose broth with 0.4 sorbitol) Washing solution 5 mM potassium phosphate (pH 4.5), 0.4 M sorbitol, 10% glycerol Electroporation solution 5 mM potassium phosphate (pH 4.5), 0.4 M sorbitol, 10% glycerol Outgrowth medium HJGS (Hogg-Jago glucose broth with 0.4 sorbitol) Cuvette 1 mm gap width Reference Buckley, N. D. et al 1999 Applied and Environmental Microbiology 65, No. 9 3800-3804 Making electrocompetent cells:

1. Cultivate cells by adding 1% overnight preculture to fresh medium. Grow cells at 37 C without agitation to a cell
density of O.D.660 of 0.5. Add glycine to a final concentration of 10% and incubate again for 1 h at 37 C. 2. Harvest by centrifugation. 3. Wash twice in ice-cold washing solutio n. 4. Resuspend in electroporation solution to a 50-fold concentration and freeze in an ethanol-dry ice bath. Store at -80 C.

Electroporation of cells:

  1. Thaw cells on ice. Add 1 g plasmid DNA to 100 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) 1,600 V Time constant (T) 5 ms
  4. Add 1 ml ice-cold outgrowth medium and incubate for 3 h at 37 C.
  5. Plate aliquots onto selective agar plates; incubate 2-3 days at 37 C.
Expected Results: Transformation efficiency up to 1 x 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Streptococcus salivarius
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/11/2016)... 11, 2016  Neurocrine Biosciences, Inc. (NASDAQ: NBIX ) today ... 31, 2015. --> --> ... loss of $29.3 million, or $0.34 loss per share, compared to ... for the same period in 2014. For the year ended December ... or $1.05 loss per share, as compared to a net loss ...
(Date:2/11/2016)... -- Wellcentive today announced it has been selected by FamilyCare ... community care organization (CCO) with more than 130,000 ... and care management solutions and services. Wellcentive,s capabilities ... managers, analysts and care managers while providing insight ... members. Oregon . ...
(Date:2/11/2016)... Buffalo, New York (PRWEB) , ... February 11, ... ... and analytical instruments for more than 150 years, continues today to pursue the ... to its line of analytical instruments: the AR9 Refractometer and the AR5 Refractometer. ...
(Date:2/10/2016)... , Feb. 10, 2016  The Maryland House of ... has announced that University of Maryland School of Medicine ... and University of Maryland Medical System President and CEO ... "Speaker,s Medallion," the highest honor given to the public ... Dean Reece and Mr. Chrencik for ...
Breaking Biology Technology:
(Date:2/9/2016)... Feb. 9, 2016 Vigilant Solutions announces today that ... (LPR) to develop a lead in a difficult homicide case. ... data to locate the suspect vehicle. Due to the ongoing ... have been omitted at the agency,s request. ... explains, "Our victim was found deceased at an intersection here ...
(Date:2/3/2016)... India , February 3, 2016 ... the new market research report "Automated Fingerprint Identification System ... Search, Latent Search), Application (Banking & Finance, Government, Healthcare, ... published by MarketsandMarkets, the market is expected to be ... CAGR of 21.0% between 2015 and 2020. The transformation ...
(Date:2/2/2016)... Feb. 2, 2016 Checkpoint Inhibitors for ... Market Are you interested in the future ... for checkpoint inhibitors. Visiongain,s report gives those predictions ... and national level. Avoid falling behind in ... opportunities and revenues those emerging cancer therapies can ...
Breaking Biology News(10 mins):