Navigation Links
Pseudomonas aeruginosa

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.528 01/2002 Microorganism Pseudomonas aeruginosa Cell type Bacteria, gram negative Molecules injected Plasmid DNA (pUC 18 with 1.8 kb insert in water) Growth medium LB medium Washing solution 300 mM sucrose Electroporation solution 300 mM sucrose Outgrowth medium LB medium Cuvette 2 mm gap width Reference Smith A.W. and Iglewski B.H. 1989 Nucleic Acids Research 17, No. 24 10509 Making electrocompetent cells:

1. Grow cells in LB medium at 37 C with shaking up to an O.D.540 of 0.3-05. 2. Harvest by centrifugation (7,000 x g, 10 min, at 4 C). 3. Wash pellet in the original volume with sucrose, centrifuge and wash again in _ volume of washing solution. 4. Resuspend in 300 mM sucrose to a final concentration of 1011 cells/ml, chill on ice for 30 minutes.

Electr oporation of cells:

  1. Add up to 5 g plasmid DNA (1 g/l) 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) 1,600 V Time constant (T) 5 ms
  4. Add 1 ml LB medium, transfer to a sterile tube containing additional 2 ml of LB. Incubate 2 hours at 37 C with
    shaking.
  5. Plate on selective LB plates.
Expected Results: Transformation efficiency up to 107 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Pseudomonas aeruginosa
2. Pseudomonas putida
3. Pseudomonas putida
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/12/2017)... ... October 12, 2017 , ... AMRI, a ... biotechnology industries to improve patient outcomes and quality of life, will now be ... are being attributed to new regulatory requirements for all new drug products, including ...
(Date:10/11/2017)... USA (PRWEB) , ... October 11, 2017 , ... ComplianceOnline’s ... take place on 7th and 8th June 2018 in San Francisco, CA. The Summit ... as well as several distinguished CEOs, board directors and government officials from around the ...
(Date:10/11/2017)... Austin, TX (PRWEB) , ... October 11, 2017 ... ... in August compared the implantation and pregnancy rates in frozen and fresh ... the contribution of progesterone and maternal age to IVF success. , After comparing ...
(Date:10/10/2017)... ... October 10, 2017 , ... Dr. Bob Harman, founder and CEO of ... Club. The event entitled “Stem Cells and Their Regenerative Powers,” was ... Dr. Harman, DVM, MPVM was joined by two human doctors: Peter B. Hanson, M.D., ...
Breaking Biology Technology:
(Date:4/11/2017)... 2017 Research and Markets has announced the ... to their offering. ... eye tracking market to grow at a CAGR of 30.37% during ... Market 2017-2021, has been prepared based on an in-depth market analysis ... and its growth prospects over the coming years. The report also ...
(Date:4/5/2017)... 5, 2017 Today HYPR Corp. , ... server component of the HYPR platform is officially ... end-to-end security architecture that empowers biometric authentication across Fortune ... already secured over 15 million users across the financial ... connected home product suites and physical access represent a ...
(Date:3/30/2017)... HONG KONG , March 30, 2017 ... developed a system for three-dimensional (3D) fingerprint identification by adopting ground ... technology into a new realm of speed and accuracy for use ... applications at an affordable cost. ... ...
Breaking Biology News(10 mins):