Navigation Links
Staphylococcus aureus

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.533 01/2002 Microorganism Staphylococcus aureus Cell type Bacteria, gram positive Molecules injected Plasmid DNA (pC194) Growth medium Basic medium (BM: 1% peptone, 0.5% yeast extract, 0.1% glucose, 0.5% NaCl,
0.1% K2HPO4) Washing solution 10% glycerol (in distilled water) Electroporation solution 10% glycerol (in distilled water) Outgrowth medium SMMP50 medium (5.5 parts SMM buffer (1 M sucrose, 0.04 M maleic acid, 0.04 M MgCl2, pH 6.5), 4 parts 7% Panassy broth, 0.5 parts 10% BSA) Cuvette 2 mm gap width Reference Augustin J. and Gtz F. 1990 FEMS Microbiology Letters 66 203-208 Making electrocompetent cells:

1. Inoculate a flask of BM with a fresh overnight culture. Grow at 37 C with shaking to an O.D.578 of about 0.5 to 0.65. 2. Wash in one volume of 10% glycerol at room temperature, followed by washes with 1/2, 1/20 and 1/50 vol umes. 3. Resuspend to final volume of 1-5 x 1010 cells/ml in 10% glycerol. Aliquote and freeze at 70 C.

Electroporation of cells:

  1. Thaw cells for 5 min at room temperature. Add 1-2 l plasmid DNA (100-500 ng) to 50 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Incubate at room temperature for 30 minutes.
  2. Transfer mixture into a cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 1,800 V Time constant (T) 5 ms
  4. Add 1 ml SMMP50 medium to the cells and transfer to sterile tube. Incubate 90 minutes at 37 C with moderate shaking.
  5. Plate on selective BM plates. Incubate 20 hours at 37 C.
Expected Results: Transformation efficiency up to 1.5 x 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Staphylococcus epidermidis
2. Staphylococcus aureus
3. Staphylococcus epidermidis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:8/16/2017)... ... August 16, 2017 , ... Tunnell Consulting announced today that ... ISPE Annual Meeting and Expo , to be held October 29 through November ... is “Driving innovation to advance patient therapies.” , The ISPE Annual Meeting and Expo ...
(Date:8/16/2017)... Linda, Ca (PRWEB) , ... August 16, 2017 ... ... treatment by changing into a different cell type. Many treatments for specific cancers, ... A prominent example of targeted treatment is androgen deprivation therapy for advanced prostate ...
(Date:8/16/2017)... ... August 16, 2017 , ... ... delivery system, announced it has secured $2M in funding from an impressive group ... Carmen Innovations, and SVG Thrive Fund. With this investment, 3Bar is broadening availability ...
(Date:8/16/2017)... WAKEFIELD, Mass. , Aug. 16, 2017  This year,s edition of ... global leader in life sciences workforce solutions, has made the list for ... Inc. 5000 recognizes the nation,s fastest-growing private companies based on a set ... Fast 50, which includes the fastest-growing companies in the Bay ... Inc. 5000 ...
Breaking Biology Technology:
(Date:4/17/2017)... , April 17, 2017 NXT-ID, Inc. (NASDAQ: ... announces the filing of its 2016 Annual Report on Form 10-K ... Commission. ... 10-K is available in the Investor Relations section of the Company,s ... the SEC,s website at http://www.sec.gov . 2016 Year ...
(Date:4/11/2017)... , Apr. 11, 2017 Research and Markets ... 2017-2021" report to their offering. ... The global eye tracking market to grow at a CAGR ... Global Eye Tracking Market 2017-2021, has been prepared based on an ... the market landscape and its growth prospects over the coming years. ...
(Date:4/5/2017)...  The Allen Institute for Cell Science today announces ... portal and dynamic digital window into the human cell. ... application of deep learning to create predictive models of ... a growing suite of powerful tools. The Allen Cell ... publicly available resources created and shared by the Allen ...
Breaking Biology News(10 mins):