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:2/4/2016)... ... 2016 , ... Shimadzu Scientific Instruments will showcase several new ... poster sessions, and present on the analysis of mycotoxins and medical cannabis at ... 10 at the Georgia World Congress Center in Atlanta, Georgia. , Attendees ...
(Date:2/4/2016)... SHENZHEN, China , Feb. 4, 2016 ... government, and various medical institutions attended a ceremony in ... provide integrative, personalized cell therapy in 2016. ... the "Shenzhen Clinical Translation Platform for Personalized Cell Therapy" ... Shenzhen Regional Cell Production Center, both subsidiaries of Beike ...
(Date:2/4/2016)... and MENLO PARK, Calif. , ... DMPI) ("DelMar" and the "Company"), a biopharmaceutical company focused on ... that it will present at the 18 th ... 8, 2016 at 10:00 a.m. EST in New ... DelMar,s president and CEO, will provide an update on the ...
(Date:2/4/2016)... , Feb. 4, 2016 ContraVir Pharmaceuticals, Inc. ... the development and commercialization of targeted antiviral therapies, announced ... Investor Conference 2016, to be held February 8-9, 2016, ... Group,s 2016 Disruptive Growth & Healthcare Conference, taking place ... 10-11, 2016. James Sapirstein , Chief Executive ...
Breaking Biology Technology:
(Date:1/22/2016)... 22, 2016 ... of the  "Global Behavioral Biometric Market ... --> http://www.researchandmarkets.com/research/4lmf2s/global_behavioral ) has announced ... Biometric Market 2016-2020"  report to their ... Markets ( http://www.researchandmarkets.com/research/4lmf2s/global_behavioral ) has announced the ...
(Date:1/20/2016)... , Jan. 20, 2016   MedNet Solutions , ... entire spectrum of clinical research, is pleased to announce ... significant achievements are the result of the company,s laser ... iMedNet eClinical , it,s comprehensive, easy-to-use and highly ... --> Key MedNet growth achievements in 2015 ...
(Date:1/13/2016)... January 13, 2016 --> ... a new market report titled - Biometric Sensors Market - ... 2015 - 2023. According to the report, the global biometric sensors ... anticipated to reach US$1,625.8 mn by 2023, expanding at ... terms of volume, the biometric sensors market is expected ...
Breaking Biology News(10 mins):