Navigation Links
Nocardia asteroides

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.525 01/2002 Microorganism Nocardia asteroides Cell type Bacteria, gram positive Molecules injected Plasmid DNA (in TE buffer) Growth medium N2 medium (20 g/l glucose, 2 g/l yeast extract, 4 g/l beef extract, 6 g/l tryptone, 2 g/l NaCl, 10 g/l glycine) Washing solution Distilled water Electroporation solution 0.3 M sucrose, 15% glycerol Outgrowth medium N2 medium (without antibiotics) Cuvette 2 mm gap width Reference Yao W., et al 1994 Current Microbiology 29 223-227 Making electrocompetent cells:

1. Inoculate a fresh overnight culture of bacteria into N2 medium. Grow cells at 28 C with shaking to an O.D.600 of 0.7. 2. Harvest by centrifugation. 3. Wash twice with _ volume of distilled water. 4. Resuspend in a final volume of 1/20 of the original volume of electroporation solution.

Electroporation of cells:

  1. Add 2 l (0.1 g) plasmid DNA to 50 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) 2,500 V Time constant (T) 5 ms
  4. Add 1 ml N2 medium and incubate for 2 hours at 30 C.
  5. Plate cells on selective YEME medium with a 3 ml YEME soft agar (0.4%) overlayment.
Expected Results: Transformation efficiency up to 8 x 104 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Nocardia asteroides
2. Nocardia corallina
3. Nocardia corallina
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:1/24/2017)... 2017   Bracket , a leading clinical trial ... of Jennifer Peters to its growing team ... Manager for the Scientific Services division. In this role, ... Rater Training, Quality Assurance, and Electronic Clinical Outcomes Assessments ... and demonstrated record building technology-based businesses in life sciences. ...
(Date:1/24/2017)... ... January 24, 2017 , ... Men ... serum leptin levels had a positive association with increased prostate growth or benign ... the International Neurourology Journal involved 571 Korean men who underwent urological examinations, ...
(Date:1/24/2017)... ... 2017 , ... The Foundation for Food and Agriculture Research, ... career nominees for the 2017 New Innovator in Food and Agriculture Research Award. ... in which one in nine people suffers from hunger. The Foundation for Food ...
(Date:1/24/2017)... 2017   Instrument Business Outlook ( ... MN ) the 2016 Company of the ... newsletter tracking developments in the analytical and life ... consistently achieved outstanding technical, operational and financial results ... of IBO. "In 2016, Bio-Techne capitalized on opportunities ...
Breaking Biology Technology:
(Date:12/19/2016)... , 19 de diciembre de 2016  Mosaic Biomedicals SL anunció ... acelerado de MSC-1, un anticuerpo humanizado que se espera comenzar a ... con múltiples sitios previstos a lo largo de Europa y Norteamérica. ... MSC-1 es el ... de leucemia (LIF), una citoquina pleiotrópica que se sobreexpresa en ciertos ...
(Date:12/15/2016)... , Dec. 15, 2016   WaferGen Bio-systems, ... held genomics technology company, announced today that on December ... Qualifications Department of The Nasdaq Stock Market LLC which ... bid price of WaferGen,s common stock had been at ... WaferGen has regained compliance with Listing Rule 5550(a)(2) of ...
(Date:12/12/2016)... Dec. 12, 2016  Researchers at Trinity College, ... graphene by combining the material with Silly Putty. The ... pressure detector able to sense pulse, blood pressure, ... spider.  The research team,s findings ... read here:  http://science.sciencemag.org/content/354/6317/1257 ...
Breaking Biology News(10 mins):