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:5/24/2017)... DC, USA (PRWEB) , ... May 23, 2017 , ... ... economy as well as an enabler of life-saving medical and other vital technologies — ... photonics . They joined others in the scientific community today in responding to the ...
(Date:5/23/2017)... ... May 23, 2017 , ... ... unlimited source of human cardiovascular cells for research and the development of ... it possible to generate large numbers of cardiomyocytes (hPSC-CMs). Due to varying ...
(Date:5/23/2017)... , ... May 23, 2017 , ... ... advanced technology applications, has announced a facility expansion to accommodate its rapid growth. ... feet of new workspace and renovation of the existing areas. The expansion includes, ...
(Date:5/22/2017)... (PRWEB) , ... May 22, 2017 , ... Stratevi, a ... expanded to the East Coast. It has opened an office in downtown Boston at ... are finding it increasingly more important to generate evidence on the value they provide, ...
Breaking Biology Technology:
(Date:5/6/2017)... RAM Group , Singaporean based ... in biometric authentication based on a novel  ... to perform biometric authentication. These new sensors are based on ... Ram Group and its partners. This sensor will have ... and security. Ram Group is a next generation ...
(Date:4/13/2017)... Calif. , April 13, 2017 UBM,s ... York will feature emerging and evolving technology ... Both Innovation Summits will run alongside the expo portion ... speaker sessions, panels and demonstrations focused on trending topics ... largest advanced design and manufacturing event will take place ...
(Date:4/11/2017)... Florida , April 11, 2017 ... a security technology company, announces the appointment of independent Directors ... Bendheim to its Board of Directors, furthering the company,s ... ... of NXT-ID, we look forward to their guidance and benefiting ...
Breaking Biology News(10 mins):