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:12/2/2016)... ... December 01, 2016 , ... Aerocom Healthcare ( http://www.aerocomhealthcare.com ... will present its chain-of-custody solution for tracking and securing medications at booth 676 ... 4-8, 2016. , Aerocom has a proven solution for tracking medications via its ...
(Date:12/2/2016)... NY (PRWEB) , ... December 01, 2016 , ... The ... is expanding to three days and will take place on February 1-3, 2017 at ... and Dr James Gulley (NCI), the program provides a unique 360-degree approach, which addresses ...
(Date:12/2/2016)... (PRWEB) , ... December 01, 2016 , ... ... congratulate long-term client Nanowear on their recent FDA Class II 510(k) clearance for ... a significant hurdle in commercializing remote cardiac monitoring devices that rely on cloth-based ...
(Date:12/2/2016)... ... December 02, 2016 , ... Robots will storm the Prudential ... December 3rd, 2016. The event, which is held on the United Nations International Day ... with Disabilities back into the workplace. Suitable Technologies is partnering with NTI to showcase ...
Breaking Biology Technology:
(Date:11/30/2016)... and WARSAW, Poland , Nov. 30, 2016 Not ... one of the most crucial aspects of recovery so we need to do it ... health risks, including heart problems, high blood pressure, stroke, diabetes, and even cancer. ... sleep and find a Christmas present that could help them to manage their sleep ... ...
(Date:11/29/2016)... BioDirection, a privately held medical device company developing novel ... concussion and other traumatic brain injury (TBI), announced today ... the U.S. Food and Drug Administration (FDA) to review ... meeting company representatives reviewed plans for clinical development of ... a planned pilot trial. "We are ...
(Date:11/29/2016)... Nearly one billion matches per second with DERMALOG,s high-speed AFIS    ... ... DERMALOG is Germany's largest Multi-Biometric supplier: The ... Identification Systems) ... Germany's largest Multi-Biometric supplier: The company's Fingerprint Identification System is part of ...
Breaking Biology News(10 mins):