Navigation Links
Corynebacterium glutamicum

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.510 12/2001 Microorganism Corynebacterium glutamicum Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium LBG (LB with 0.5% glucose) supplemented with 2.5% glycine and isonicotinic acid hydrazid (4 mg/ml) Washing solution 15% glycerol (v/v) Electroporation solution 15% glycerol (v/v) Outgrowth medium LBG medium (LB with 0.5% glucose) Cuvette 2 mm gap width Reference Haynes J. and Britz M. 1990 Journal of General Microbiology 136 255-263 Making electrocompetent cells:

1. Grow cells in LBG medium at 30 C and shaking at 200 rpm to an O.D.600 of 0.15 to 0.25. 2. Harvest by centrifugation. 3. Wash in one culture volume of 15% glycerol. 4. Resuspend cells in 0.002 culture volumes of 15% glycerol (number of cells: approx. 2.5 x 1010 cells/ml).

Electroporation of cells:

  1. Add 1-2 l plasmid DNA (in water) to a minimum of 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) 2,500 V Time constant (T) 5 ms
  4. Immediately transfer cell suspension into LBG medium at a 1:25 dilution, incubate 1 hour at 30 C.
  5. Plate cells on selective plates; incubate for 4 days.
Expected Results: Transformation efficiency up to 5 x 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Corynebacterium glutamicum
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:4/19/2017)... , ... April 18, 2017 , ... A number of ... This webinar, which is part of the Protein and Cell Analysis Education Webinar ... outline where this technology fits in current and future applications. , Many flow ...
(Date:4/19/2017)... ... April 19, 2017 , ... Nobilis Therapeutics Announces Completion of Landmark ... Clinical Data in its Upcoming Post Traumatic Stress Disorder Trial , Nobilis Therapeutics, ... trial assessing efficacy of its NBTX-001, a xenon-based therapeutic in the treatment of Panic ...
(Date:4/19/2017)... ... April 19, 2017 , ... As part of the ... to novices as well as experienced users, attendees will gain a better understanding ... tests. , Hemostasis testing quality is determined by preanalytical variables which encompass ...
(Date:4/19/2017)... and WESTMINSTER, Colo. ... LLC ("Oxford"), an industry-leading specialty finance firm that ... services companies, today announced the closing of a ... Inc. ("Cerapedics") a privately-held orthobiologics company engaged in ... products for the treatment of orthopedic injuries. ...
Breaking Biology Technology:
(Date:4/11/2017)... , Apr. 11, 2017 Research and ... Market 2017-2021" report to their offering. ... The global eye tracking market to grow at a ... report, Global Eye Tracking Market 2017-2021, has been prepared based on ... covers the market landscape and its growth prospects over the coming ...
(Date:4/5/2017)... , April 5, 2017  The Allen Institute ... Allen Cell Explorer: a one-of-a-kind portal and dynamic digital ... 3D imaging data, the first application of deep learning ... human stem cell lines and a growing suite of ... platform for these and future publicly available resources created ...
(Date:4/3/2017)... , April 3, 2017  Data ... precision engineering platform, detected a statistically significant ... product prior to treatment and objective response ... the potential to predict whether cancer patients ... to treatment, as well as to improve ...
Breaking Biology News(10 mins):