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:8/15/2017)... ... August 15, 2017 , ... ... that accelerate pharmaceutical and biotherapeutics development, announces the launch of the new ... needed to gain kinetic binding data for a wide range of molecules, including ...
(Date:8/15/2017)... ... August 15, 2017 , ... Any expert in stem cell research or stem ... more than half a century. Despite their essential roles in human health and ... that molecular tags developed for this purpose also tag other, more abundant, non-stem tissue ...
(Date:8/15/2017)... ... , ... The Conference Forum and The Trout Group ... of upcoming panels and events. The partnership culminates with the 4th Annual IO360° ... York City. , “With our experience in producing the Immuno-Oncology 360° NYC event and ...
(Date:8/15/2017)... ... 2017 , ... JULABO USA introduces its new website ... it easy to navigate through the site whether you’re in the office, in ... educational industry content and visit the company’s social media accounts, all on one ...
Breaking Biology Technology:
(Date:5/16/2017)... TEANECK, N.J. , May 16, 2017  Veratad ... leading provider of online age and identity verification solutions, ... the K(NO)W Identity Conference 2017, May 15 thru May ... Ronald Regan Building and International Trade Center. ... across the globe and in today,s quickly evolving digital ...
(Date:4/24/2017)... , April 24, 2017 ... and partner with  Identity Strategy Partners, LLP (IdSP) ... "With or without President Trump,s March 6, 2017 ... Terrorist Entry , refugee vetting can be instilled with ... resettlement. (Right now, all refugee applications are suspended ...
(Date:4/17/2017)... NXT-ID, Inc. (NASDAQ: NXTD ) ("NXT-ID" or the "Company"), ... Report on Form 10-K on Thursday April 13, 2017 with the ... The ... section of the Company,s website at http://www.nxt-id.com  under "SEC Filings," ... 2016 Year Highlights: Acquisition of ...
Breaking Biology News(10 mins):