Navigation Links
Listeria monocytogenes

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.520 12/2001 Microorganism Listeria monocytogenes 23074 Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium BHI, supplemented with 0.5 M sucrose Washing solution 1 mM HEPES (pH 7.0), 0.5 M sucrose Electroporation solution 1 mM HEPES (pH 7.0), 0.5 M sucrose Outgrowth medium BHI, supplemented with 0.5 M sucrose Cuvette 1 mm gap width Reference Park, S. and Stewart, G. 1990 Gene 94 129-132 Making electrocompetent cells:

1. Dilute an overnight culture of L. monocytogenes 23074 in BHI, into fresh media (1:100). Grow at 37 C with shaking until reaching an O.D.600 of 0.2. 2. Add Penicillin G (10 g/ml) and continue incubation for a further 2 hours. 3. Harvest by centrifugation (8,000 x g, 10 min., at 4 C) and wash three times in ice-cold washing solution, once with equal volume and twice with _ volume. 4. Resuspend the cells in ice-cold electroporation solution (0.0025 vol.). Use cells within 30 minutes of their preparation.

Electroporation of cells:

  1. Add 1 g plasmid DNA to 100 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) 1,000 V Time constant (T) 5 ms
  4. Immediately add 1 ml of BHI medium and incubate at 37 C for one hour.
  5. Plate cells on selective BHI plates.
Expected Results: Transformation efficiency up to 4 x 106 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Listeria monocytogenes
2. Two-dimensional Optimization of a Semi-nested PCR for Detecting Listeria monocytogenes
3. Use of the DCode System to Detect the Food-Borne Bacterial Pathogen Listeria monocytogenes
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:3/30/2017)... ... March 30, 2017 , ... ... four posters at the American Association for Cancer Research (AACR) Annual Meeting ... of Vortex’s technology to rapidly collect highly enriched populations of CTCs, undamaged by ...
(Date:3/29/2017)... ... March 29, 2017 , ... Zansors announced today ... as issued by the U.S. Patent & Trademark Office (USPTO). The patent covers ... and health monitoring. This invention will be critical to the future of wearable ...
(Date:3/29/2017)... -- "Surging application of gesture control features in automobiles ... to drive the growth of gesture recognition and touchless ... expected to be worth USD 18.98 billion by 2022, ... 2022. The touchless sensing market is expected to be ... CAGR of 17.44% between 2017 and 2022. Gesture recognition ...
(Date:3/29/2017)... Palo Alto, CA, USA (PRWEB) , ... March 29, 2017 , ... ... experts in various regulatory subjects has announced its 3rd Annual Medical Device Summit 2017 ... 2017 at the Omni Parker House Hotel, in Boston, MA. , The Omni Parker ...
Breaking Biology Technology:
(Date:2/26/2017)... DALLAS , Feb. 25, 2017  Securus ... justice technology solutions for public safety, investigation, corrections ... of Recidivism and Reentry. "Too often, ... State prisons and county jails are trying to ... of inmates and friends and family members. While significant ...
(Date:2/22/2017)... , Feb. 22, 2017 With ... 2021, ABI Research identifies four technologies that innovative ... to secure significant share in the changing competitive ... and passive authentication.   "Companies can ... comes to security," says Dimitrios Pavlakis , ...
(Date:2/21/2017)... , February 21, 2017 ... 70 Millionen US-Dollar wachsen. Nach einem Gespräch mit mehr als ... einige Hindernisse zu überwinden gilt, um diese Prognose zu ... ... anderem die Mobilisierung der finanziellen Mittel für die Biobank, ...
Breaking Biology News(10 mins):