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.



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:

(Date:6/24/2016)... TOKYO , June 24, 2016  Regular discussions on ... to take place between the two entities said Poloz. ... in Ottawa , he pointed to the ... and the federal government. ... Poloz said, "Both institutions have common economic goals, why not ...
(Date:6/24/2016)... ... June 24, 2016 , ... While the majority of commercial spectrophotometers and ... and the 6000i models are higher end machines that use the more unconventional z-dimension ... light beam from the bottom of the cuvette holder. , FireflySci has developed ...
(Date:6/23/2016)... WA (PRWEB) , ... June 23, 2016 , ... ... announces the release of its second eBook, “Clinical Trials Patient Recruitment and Retention ... recruitment and retention in this eBook by providing practical tips, tools, and strategies ...
(Date:6/23/2016)... , June 23, 2016   Boston ... of novel compounds designed to target cancer stemness ... has been granted Orphan Drug Designation from the ... treatment of gastric cancer, including gastroesophageal junction (GEJ) ... inhibitor designed to inhibit cancer stemness pathways by ...
Breaking Biology Technology:
(Date:6/16/2016)... June 16, 2016 The ... expected to reach USD 1.83 billion by 2024, ... Research, Inc. Technological proliferation and increasing demand in ... expected to drive the market growth. ... The development of advanced multimodal techniques for ...
(Date:6/3/2016)... , June 3, 2016 ... von Nepal hat ... Lieferung hochsicherer geprägter Kennzeichen, einschließlich Personalisierung, Registrierung ... in der Produktion und Implementierung von Identitätsmanagementlösungen. ... Ausschreibung im Januar teilgenommen, aber Decatur wurde ...
(Date:5/24/2016)... Ampronix facilitates superior patient care by providing unparalleled technology to leaders of ... the latest premium product recently added to the range of products distributed by Ampronix. ... ... ... Medical Display- Ampronix News ...
Breaking Biology News(10 mins):