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:12/5/2016)... ... , ... In anticipation of AxioMed’s exclusive cleanroom manufacturing facility ... Jake Lubinski will be traveling to Germany on December 6th. Mr. Lubinski will ... to discuss the benefits of a viscoelastic total disc replacement. , AxioMed received ...
(Date:12/5/2016)... /PRNewswire/ - Resverlogix Corp. ("Resverlogix" or the "Company") ... Safety Monitoring Board (DSMB) for the Company,s Phase ... patients has completed a second planned safety review ... planned without any modifications. The DSMB reviewed available ... efficacy concerns were identified. The DSMB will conduct ...
(Date:12/5/2016)... Research and Markets has announced the addition of Jain ... Companies" to their offering. ... , , ... development of sequencing technologies, and their applications. Current large and ... Various applications of sequencing are described including those for genetics, ...
(Date:12/4/2016)... ... December 03, 2016 , ... ... studies. A microbiome impact grant award has been made to Dr. Renato Polimanti ... smoking and drinking on the oral microbiome. Grant proposals have been vetted by ...
Breaking Biology Technology:
(Date:11/29/2016)... Nov. 29, 2016 BioDirection, a privately held ... for the objective detection of concussion and other traumatic ... successfully completed a meeting with the U.S. Food and ... test Pre-Submission Package. During the meeting company representatives reviewed ... a precursor to commencement of a planned pilot trial. ...
(Date:11/22/2016)... According to the new market research report "Biometric ... Signature, Voice), Multi-Factor), Component (Hardware and Software), Function (Contact and Non-contact), Application, ... is expected to grow from USD 10.74 Billion in 2015 to reach ... and 2022. Continue Reading ... ...
(Date:11/17/2016)... , Nov. 17, 2016  AIC announces that it has just released a ... organizations that require high-performance scale-out plus high speed data transfer storage solutions. ... ... ... Setting up a high performance ...
Breaking Biology News(10 mins):