Navigation Links
Pseudomonas putida

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.529 01/2002 Microorganism Pseudomonas putida P8 Cell type Bacteria, gram negative Molecules injected Plasmid DNA Growth medium Standard 1-medium Washing solution 10% glycerol Electroporation solution 10% glycerol Outgrowth medium Standard 1-medium Cuvette 2 mm gap width Reference Prof. Friedhelm Meinhardt Universitt Mnster Corrensstrae 3 D-48149 Mnster
e-mail: meinhar@uni-muenster.de Making electrocompetent cells:

1. Inoculate 50 ml standard-1 medium with 7 ml of a fresh overnight culture of Pseudomonas putida. Grow cells at 30 C to a density of O.D. of 0.8. 2. Harvest by centrifugation. 3. Wash twice with 50 ml ice-cold glycerol, centrifuge. 4. Resuspend cells in 0.8 ml ice-cold glycerol, keep on ice.

Electroporation of cells:

  1. Add 4 l plasmid DNA (1 g) 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) 2,400 V Time constant (T) 5 ms
  4. Immediately add 1 ml standard 1-medium. Incubate 2 hours at 30 C.
  5. Plate cells on selective plates.
Expected Results: Transformation efficiency up to 2 x 104 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

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


(Date:10/11/2017)... BioMarketing, a leading provider of patient support solutions, has announced ... network, which will launch this week. The VMS CNEs will ... to enhance the patient care experience by delivering peer-to-peer education ... professionals to help women who have been diagnosed and are ... ...
(Date:10/11/2017)... ... October 11, 2017 , ... Singh Biotechnology today announced ... to SBT-100, its novel anti-STAT3 (Signal Transducer and Activator of Transcription 3) B ... to cross the cell membrane and bind intracellular STAT3 and inhibit its function. ...
(Date:10/10/2017)... , ... October 10, 2017 , ... Dr. Bob Harman, ... his local San Diego Rotary Club. The event entitled “Stem Cells ... and had 300+ attendees. Dr. Harman, DVM, MPVM was joined by two human ...
(Date:10/10/2017)... ... October 10, 2017 , ... The Pittcon Program Committee ... honoring scientists who have made outstanding contributions to analytical chemistry and applied ... the world’s leading conference and exposition for laboratory science, which will be held ...
Breaking Biology Technology:
(Date:4/5/2017)... -- The Allen Institute for Cell Science today announces the ... and dynamic digital window into the human cell. The ... of deep learning to create predictive models of cell ... growing suite of powerful tools. The Allen Cell Explorer ... available resources created and shared by the Allen Institute ...
(Date:3/30/2017)... -- On April 6-7, 2017, Sequencing.com will host the world,s ... at Microsoft,s headquarters in Redmond, Washington ... health and wellness apps that provide a unique, personalized ... is the first hackathon for personal genomics and the ... the genomics, tech and health industries are sending teams ...
(Date:3/27/2017)... , March 27, 2017  Catholic Health ... and Management Systems Society (HIMSS) Analytics for achieving ... Adoption Model sm . In addition, CHS previously ... U.S. hospitals using an electronic medical record (EMR). ... its high level of EMR usage in an ...
Breaking Biology News(10 mins):