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.



Page: All 1 2

Related biology technology :

1. Agrobacterium tumefaciens
Post Your Comments:

(Date:10/13/2015)... -- PharmaCyte Biotech, Inc. (OTCQB: PMCB) shares closed at $.1035, ... average with 1,308,352 shares being traded. PMCB shares have been ... low of $.072 in September, then bouncing back to current ... due diligence will show investors in PharmaCyte Biotech, Inc. information ... using the link below at no cost. ...
(Date:10/13/2015)... ... October 13, 2015 , ... ... and assessment of organotypic 3D cell culture models, has launched a 14 ... in their patent-pending 3D InSightâ„¢ Human Liver Microtissues. The service streamlines toxicity ...
(Date:10/13/2015)... , ... October 13, 2015 , ... ... President. Steve joins as the Pistoia Alliance continues to drive to increase the ... non-profit Pistoia Alliance is focusing its vision and expanding its work in supporting ...
(Date:10/13/2015)... Oct. 13 2015 Research and Markets( ... & Europe Markets for Bone Morphogenetic Protein Growth Factor ... offering. --> --> ... the formation of bone after a fracture. In nature, ... in the formation of the skeleton. There are twenty ...
Breaking Biology Technology:
(Date:9/28/2015)... 2015 Synaptics Inc. (NASDAQ: SYNA ), ... that Lenovo has selected Synaptics , Natural ID ... latest smartphone, the Vibe P1. The new Vibe ... device and provide swift access to applications and mobile ... ID FS4202 sensor solution utilizes AES256-bit encryption of the ...
(Date:9/24/2015)... , Sept. 24, 2015  EyeLock LLC, a ... showcasing its award winning and latest technology in Booth ... Anaheim, California . EyeLock,s iris authentication ... with unmatched biometric accuracy, making it the most proven ... platform uses video technology to deliver a fast and ...
(Date:9/10/2015)... This report provides detailed descriptions of the sensor types ... types that will dominate in the future. Many product ... technology hype curve in the last five years before ... all of them is the prominence of sensor options ... Sensors collect data about the physical and chemical properties ...
Breaking Biology News(10 mins):