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:2/8/2016)... ... , ... Bulk food product inspection systems are specifically designed ... the production process. Despite frequently inspecting loose product prior to packaging, product inspection ... sacks of dry powders. , Mettler-Toledo Product Inspection's brand-new white paper entitled "Improving ...
(Date:2/8/2016)... the development of future natural products for crop protection. In ... center in Israel and invests substantial ... has a variety of products adapted to regional needs, ... than 35 counties worldwide. ... Gold ® is used to control a broad spectrum ...
(Date:2/6/2016)... McLean, VA (PRWEB) , ... February 06, 2016 , ... ... enrichment session, cost-free, for middle and high school teachers on Wednesday February 10, 2016. ... be held at the Smithsonian-Mason School of Conservation, located at 1500 Remount Road in ...
(Date:2/5/2016)... , Feb. 5, 2016  In the pharmaceutical industry ... a host of launch activities including the identification and ... launch activity is especially high in the oncology therapeutic ... Best Practices and the Role of Medical Affairs in ... focused on oncology therapies find better ways to utilize ...
Breaking Biology Technology:
(Date:1/13/2016)... 2016 http://www.researchandmarkets.com/research/7h6hnn/india_biometrics ... the  "India Biometrics Authentication & Identification ...  report to their offering.  ... the addition of the  "India Biometrics ... & Forecast (2015-2020)"  report to ...
(Date:1/8/2016)... MANCHESTER, United Kingdom , Jan. 8, 2016 /PRNewswire/ ... sensor-based diagnostic products, today announced the closing of a $9 ... investors.  Proceeds from the financing will be used to accelerate ... device for detecting early-stage pressure ulcers. ... after receiving CE Mark approval. The device,s introduction has been ...
(Date:1/7/2016)... 2016 Various factors have contributed to ... as biologics and biosimilars. Some of these factors ... growing demand for cost-effective alternatives, growing burden of ... similar versions of their corresponding patented biologic drugs, ... safety, and efficacy. The global biosimilars market is ...
Breaking Biology News(10 mins):