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:2/8/2016)... , Feb. 8, 2016 Novan, Inc. today announced ... of the Board of Directors of Novan. In addition, Robert ... North Carolina . --> ... also announced that it received a total of $32.8 million of ... from its private investor network originating throughout the Research Triangle area ...
(Date:2/8/2016)... ... February 08, 2016 , ... Franz Inc. ... Graph Database technology, today announced the availability of AllegroGraph 6, the leading Semantic ... Cloudera Certified Technology Program (CCPT). AllegroGraph is the first Semantic Graph ...
(Date:2/6/2016)... ... ... The Center for Excellence in Education (CEE) will sponsor a Bite of ... 10, 2016. This Bite of Science session, hosted by the Smithsonian-Mason School of ... Road in Front Royal, VA from 5:00 p.m. to 8:00 p.m. The dinner is ...
(Date:2/5/2016)... , February 5, 2016 Amarantus ... biotechnology company focused on developing products for Regenerative Medicine, ... Rare Pediatric Disease Designation (RPDD) from the US Food ... with MANF. MANF was previously granted orphan drug designation ... --> Amarantus BioScience Holdings, Inc. (OTCQB: ...
Breaking Biology Technology:
(Date:2/3/2016)... -- --> --> Fourth quarter 2015: ... 1,187% compared with fourth quarter of 2014. Gross margin was ... 30.0). Earnings per share increased to SEK 6.39 (loss: 0.49). ... 74.7). , --> --> ... M (233.6), up 1,142% compared with 2014. Gross margin was ...
(Date:2/2/2016)... This BCC Research report provides a ... the recent advances in high throughput ‘omic platforms ... forward. Includes forecast through 2019. Use ... opportunities that exist in the bioinformatic market. Analyze ... well as IT and bioinformatics service providers. Analyze ...
(Date:2/1/2016)... Rising sales of consumer electronics ... intuitive gesture control market size ... consumer electronics coupled with new technological advancements to drive ... through 2020   --> ... advancements to drive global touchfree intuitive gesture control market ...
Breaking Biology News(10 mins):