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:5/24/2017)... ... May 24, 2017 , ... Today, the South ... Recovery’s Advanced Biological Nutrient Recovery (ABNR™) technology at its 4,000,000 gallon per day ... upgrade to sustainably meet current and future nutrient discharge regulations. The ABNR platform, ...
(Date:5/24/2017)... May 24, 2017 (PRWEB) , ... May 24, ... ... Connectivity , Medical systems are increasingly being developed with Wi-Fi connectivity to reduce ... easily moved from room to room. In addition, compact mobile devices including infusion ...
(Date:5/23/2017)... ... May 23, 2017 , ... A recent survey conducted by the Weed ... to control weed in 12 categories of broadleaf crops, fruits and vegetables, while common ... the U.S. and Canada participated in the 2016 survey, the second conducted by WSSA. ...
(Date:5/23/2017)... ... May 22, 2017 , ... NetDimensions has ... the Aragon Research Globe™ for Corporate Learning, 2017. , Aragon Research defines Leaders ... demand, and effectively perform against those strategies. NetDimensions’ ranking as a Leader due ...
Breaking Biology Technology:
(Date:3/30/2017)... March 30, 2017  On April 6-7, 2017, Sequencing.com ... Genome hackathon at Microsoft,s headquarters in ... will focus on developing health and wellness apps that ... Hack the Genome is the first hackathon for ... world,s largest companies in the genomics, tech and health ...
(Date:3/30/2017)... NEW YORK , March 30, 2017 ... by type (physiological and behavioral), by technology (fingerprint, AFIS, ... recognition, voice recognition, and others), by end use industry ... travel and immigration, financial and banking, and others), and ... Europe , Asia Pacific ...
(Date:3/28/2017)... 2017 The report "Video Surveillance ... Servers, Storage Devices), Software (Video Analytics, VMS), and Service ... Forecast to 2022", published by MarketsandMarkets, the market was ... projected to reach USD 75.64 Billion by 2022, at ... base year considered for the study is 2016 and ...
Breaking Biology News(10 mins):