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


(Date:3/23/2017)... ... March 23, 2017 , ... ... exhibits both viscous and elastic characteristics when deformed, which is identical to how ... to gently absorb compressive forces and return to its natural state along a ...
(Date:3/23/2017)... 23, 2017  Northwest Biotherapeutics (OTCQB: NWBO) (NW ... therapies for solid tumor cancers, today announced that ... it announced last Friday, March 17, 2017. ... investors securities totaling 28,843,692 shares, comprised of 18,843,692 ... shares of Class C Warrants pre-funded at the ...
(Date:3/23/2017)... Calif., March 23, 2017  BioPharmX Corporation (NYSE ... for the dermatology market, today reported financial results ... 2017, and will provide an update on the ... year. "We are pleased to report ... for BioPharmX," said President Anja Krammer. "We achieved ...
(Date:3/23/2017)... -- NetworkNewsWire Editorial Coverage  ... Cancer remains one of the ... care systems, in terms of costs and resources. However, as ... development of innovative and efficient therapies that demonstrate higher chances ... of cancer treatments, a growing number of patients receiving immuno-oncology ...
Breaking Biology Technology:
(Date:2/28/2017)... February 28, 2017 News solutions for biometrics, ... ... from 14 to 16 March, Materna will present its ... how seamless travel is a real benefit for passengers. To ... to their passenger touch point solutions to take passengers through the ...
(Date:2/22/2017)... LONDON , Feb. 22, 2017 ... by 2021, ABI Research identifies four technologies that ... solutions to secure significant share in the changing ... payments, and passive authentication.   "Companies ... it comes to security," says Dimitrios Pavlakis ...
(Date:2/13/2017)... FRANCISCO , Feb. 13, 2017  RSA ... centralized platform that is designed to enhance fraud ... latest release in the RSA Fraud & Risk ... enable organizations to leverage additional insights from internal ... tools to better protect their customers from targeted ...
Breaking Biology News(10 mins):