Navigation Links
Agrobacterium tumefaciens

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.502 12/2001 Microorganism Agrobacterium tumefaciens Cell type Bacteria, gram negative Molecules injected Plasmid DNA (in water) Growth medium TB medium Washing solution Sterile, cold water Electroporation solution 10% glycerol Outgrowth medium LB medium Cuvette 1 mm gap width Reference Mersereau M., et al 1990 Gene 90 149-151

Making electrocompetent cells:

  1. Grow cells at 30 C to a density of O.D.600: 1.5-2.0.

  2. Harvest by centrifugation and wash five to seven times in sterile, cold water.

  3. Resuspend the cells in 10% glycerol to a final concentration of 4-6 x 1011 cell/ml and store frozen at 70 C.

  4. For electroporation frozen cells were thawed in ice and diluted with sterile water to a density of 1 x 1010 cells/ml prior to use.

Electroporation of cells:

  1. Add 2 ng plasmid DNA to 40 l of electrocompetent cells. Homogenize by gently mixing with pipette several times.

  2. Transfer mixture into a prechilled cuvette.
  3. Wipe moisture from the cuvette and insert the cuvette into the device.

  4. Electroporation:


  5. Mode Prokaryotes O Voltage (V) 1440 V Time constant (T) 5 ms
  6. Immediately add 400 l LB medium and transfer into a sterile tube. Incubate at room temperature for 1 hour.
  7. Plate cells on minimal selective AB plates; incubate 12-72 hours.

Expected results:

Transformation efficiency up to 1-3 x 108 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Agrobacterium tumefaciens
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/27/2016)... , June 27, 2016  Sequenom, Inc. (NASDAQ: ... enabling healthier lives through the development of innovative products ... the United States denied its ... the claims of Sequenom,s U.S. Patent No. 6,258,540 (",540 ... criteria established by the Supreme Court,s Mayo Collaborative Services ...
(Date:6/27/2016)... ... June 27, 2016 , ... Rolf K. Hoffmann, ... faculty of the University of North Carolina Kenan-Flagler Business School effective ... at UNC Kenan-Flagler, with a focus on the school’s international efforts, leading classes ...
(Date:6/24/2016)... June 24, 2016 Epic Sciences unveiled ... cancers susceptible to PARP inhibitors by targeting homologous ... (CTCs). The new test has already been incorporated ... multiple cancer types. Over 230 clinical ... response pathways, including PARP, ATM, ATR, DNA-PK and ...
(Date:6/23/2016)... ... June 23, 2016 , ... UAS LifeSciences, one of ... their brand, UP4™ Probiotics, into Target stores nationwide. The company, which has been ... Target to its list of well-respected retailers. This list includes such fine stores ...
Breaking Biology Technology:
(Date:3/31/2016)... , March 31, 2016   ... ("LegacyXChange" or the "Company") LegacyXChange is excited ... of its soon to be launched online site for ... https://www.youtube.com/channel/UCyTLBzmZogV1y2D6bDkBX5g ) will also provide potential shareholders a ... DNA technology to an industry that is notorious for ...
(Date:3/23/2016)... Massachusetts , March 23, 2016 /PRNewswire/ ... im Interesse erhöhter Sicherheit Gesichts- und Stimmerkennung ... Xura, Inc. (NASDAQ: MESG ), ... bekannt, dass das Unternehmen mit SpeechPro zusammenarbeitet, ... aus der Finanzdienstleistungsbranche, wird die Möglichkeit angeboten, ...
(Date:3/21/2016)... March 22, 2016 Unique ... passcodes for superior security   ... provider of secure digital communications services, today announced it ... and offer enterprise customers, particularly those in the Financial ... and voice authentication within a mobile app, alongside, and ...
Breaking Biology News(10 mins):