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:10/12/2017)... York, NY (PRWEB) , ... October 12, 2017 ... ... Academy of Sciences today announced the three Winners and six Finalists of the ... are given annually by the Blavatnik Family Foundation and administered by the New ...
(Date:10/12/2017)... ... 12, 2017 , ... AMRI, a global contract research, development ... patient outcomes and quality of life, will now be offering its impurity solutions ... new regulatory requirements for all new drug products, including the finalization of ICH ...
(Date:10/11/2017)... (PRWEB) , ... October 11, 2017 , ... Personal eye wash is a basic first ... eye at a time. So which eye do you rinse first if a dangerous substance ... Plum Duo Eye Wash with its unique dual eye piece. , “Whether its ...
(Date:10/11/2017)... ROTTERDAM, the Netherlands and LAGUNA HILLS, ... that The Institute of Cancer Research, London ... will use MMprofilerâ„¢ with SKY92, SkylineDx,s prognostic tool to risk-stratify ... high-risk trial known as MUK nine . The University ... this trial, which is partly funded by Myeloma UK, and ...
Breaking Biology Technology:
(Date:5/6/2017)... , May 5, 2017 ... just announced a new breakthrough in biometric authentication ... exploits quantum mechanical properties to perform biometric authentication. These ... smart semiconductor material created by Ram Group and ... finance, entertainment, transportation, supply chains and security. Ram ...
(Date:4/19/2017)... The global military biometrics market ... by the presence of several large global players. The ... major players - 3M Cogent, NEC Corporation, M2SYS Technology, ... 61% of the global military biometric market in 2016. ... military biometrics market boast global presence, which has catapulted ...
(Date:4/17/2017)... 2017 NXT-ID, Inc. (NASDAQ: NXTD ) ... of its 2016 Annual Report on Form 10-K on Thursday April ... ... in the Investor Relations section of the Company,s website at ... at http://www.sec.gov . 2016 Year Highlights: ...
Breaking Biology News(10 mins):