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:6/27/2016)... CLEVELAND , June 27, 2016  Global ... average 4.6 percent through 2020 to $7.2 billion.  ... (food and beverages, cleaning products, biofuel production, animal ... and biotechnology, diagnostics, and biocatalysts). Food and beverages ... gains driven by increasing consumption of products containing ...
(Date:6/27/2016)... ... 2016 , ... Parallel 6 , the leading software as a service ... Virtual Patient Encounter CONSULT module which enables both audio and video telemedicine communication ... , Using the CONSULT module, patients and physicians can schedule a face to face ...
(Date:6/27/2016)... , June 27, 2016   Ginkgo Bioworks ... to industrial engineering, was today awarded as one ... selection of the world,s most innovative companies. Ginkgo ... scale for the real world in the nutrition, ... engineers work directly with customers including Fortune 500 ...
(Date:6/24/2016)... ... 2016 , ... Researchers at the Universita Politecnica delle Marche in Ancona combed ... pleural mesothelioma. Their findings are the subject of a new article on the Surviving ... signposts in the blood, lung fluid or tissue of mesothelioma patients that can help ...
Breaking Biology Technology:
(Date:5/24/2016)... facilitates superior patient care by providing unparalleled technology to leaders of the medical imaging ... product recently added to the range of products distributed by Ampronix. Photo ... ... ... News ...
(Date:5/3/2016)... 2016  Neurotechnology, a provider of high-precision biometric ... Biometric Identification System (ABIS) , a complete system ... ABIS can process multiple complex biometric transactions with ... fingerprint, face or iris biometrics. It leverages the ... MegaMatcher Accelerator , which have been used ...
(Date:4/19/2016)... 2016 The new GEZE SecuLogic ... web-based "all-in-one" system solution for all door components. It ... the door interface with integration authorization management system, and ... The minimal dimensions of the access control and the ... installations offer considerable freedom of design with regard to ...
Breaking Biology News(10 mins):