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:1/17/2017)... , ... January 17, 2017 , ... ... re-engineered Drug Safety Technology Consortium™ (SafeTEC™), $3 million in investment towards 15+ TEC ... new tools and assays, and their applicability in drug safety assessment, for the ...
(Date:1/17/2017)... , ... January 17, 2017 , ... ... and epigenetics research, recently announced a collaboration with the Heidelberg University Hospital and ... method for library preparation, following the company’s successful launch of its CATS ...
(Date:1/17/2017)... -- On January 10 at the Medtech Showcase held in ... in San Francisco , ProclaRx CEO, ... pharmaceutical leaders and public and private investors about the ... destroy biofilms.  Biofilms are a physical ... body,s immune system from eradicating chronic infections. Infections with biofilms ...
(Date:1/16/2017)... MANHASSET, N.Y. , Jan. 16, 2017   ... Kevin J. Tracey, MD , president and CEO of ... Northwell Health, completed an analysis of how the nervous ... further identify and develop bioelectronic medicine devices ... today in Nature Neuroscience . The ...
Breaking Biology Technology:
(Date:12/15/2016)... 2016 "Increase in mobile transactions is driving ... biometrics market is expected to grow from USD 4.03 ... at a CAGR of 29.3% between 2016 and 2022. ... growing demand for smart devices, government initiatives, and increasing ... component is expected to grow at a high rate ...
(Date:12/7/2016)... BioCatch , the global leader in behavioral biometrics, reported a ... over 40 granted and pending patents. ... , The Company,s IP portfolio expansion ... Device, and Method Estimating Force Applied to a Touch Surface, " ... components needed to estimate the force and pressure applied to touch surfaces ...
(Date:12/6/2016)... Dec. 6, 2016 Securus Technologies, a ... solutions for public safety, investigation, corrections and monitoring, ... today a five (5) year funding commitment by ... expand the rehabilitation and reentry support to more ... Established in 2004, the Prison Entrepreneurship Program (PEP) ...
Breaking Biology News(10 mins):