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.

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


(Date:12/7/2016)...  Vyriad Inc. announced today the appointment of ... Directors. "We are delighted to welcome Al ... develop our oncolytic viruses as the next generation of ... , MD, PhD, CEO of Vyriad. "Al and his ... passion for making a difference for cancer patients—their philanthropy ...
(Date:12/7/2016)... Texas , Dec. 7, 2016  Genprex, ... of revolutionary immunogene therapy treatments, today announced that ... practice within a leading strategic communications and advisory ... communications program. The program will combine investor relations, ... objective of raising the profile of Genprex and ...
(Date:12/7/2016)... BOULDER, Colo. , Dec. 7, 2016  Muse ... genome engineering technologies, today announced that Dr. Kevin ... member of the Board of Directors. ... , who becomes the company,s Chief Science Officer as ... and Executive Director of the BioDesign Center at the ...
(Date:12/7/2016)... ANN ARBOR, Mich. , Dec. 7, 2016 /PRNewswire/ ... immune modulatory medicines, announced today the initiation of a ... candidate, LYC-30937- E nteric C oated, in patients ... that is estimated to affect as many as 7.5 million ... with approximately 1.5 - 3 million cases being diagnosed ...
Breaking Biology Technology:
(Date:11/19/2016)... 18, 2016 Securus Technologies, a leading provider ... safety, investigation, corrections and monitoring, announced today that it ... to have an independent technology judge determine who has ... high tech/sophisticated telephone calling platform, and the best customer ... do most of what we do – which clearly ...
(Date:11/17/2016)... , Nov. 17, 2016  AIC announces that it has just released a new ... that require high-performance scale-out plus high speed data transfer storage solutions. Photo ... ... ... Setting up a high performance computing ...
(Date:11/15/2016)... , Nov. 15, 2016  Synthetic Biologics, Inc. ... therapeutics focused on the gut microbiome, today announced ... 25,000,000 shares of its common stock and warrants ... at a price to the public of $1.00 ... Synthetic Biologics from the offering, excluding the proceeds, ...
Breaking Biology News(10 mins):