Navigation Links
Staphylococcus epidermidis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.534 01/2002 Microorganism Staphylococcus epidermidis 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 volumes. 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 SMMP 50 medium to the cells and transfer to sterile tube. Incubate 90 minutes at 37 C with moderate
  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.



Page: All 1 2

Related biology technology :

1. Staphylococcus aureus
2. Staphylococcus aureus
3. Staphylococcus epidermidis
Post Your Comments:

(Date:6/23/2016)... June 23, 2016   EpiBiome , a precision ... million in debt financing from Silicon Valley Bank (SVB). ... and to advance its drug development efforts, as well ... "SVB has been an incredible strategic partner ... a traditional bank would provide," said Dr. Aeron ...
(Date:6/23/2016)... ... ... STACS DNA Inc., the sample tracking software company, today announced that Dr. ... STACS DNA as a Field Application Specialist. , “I am thrilled that Dr. ... STACS DNA. “In further expanding our capacity as a scientific integrator, Hays brings a ...
(Date:6/23/2016)... 2016 Apellis Pharmaceuticals, Inc. today announced ... of its complement C3 inhibitor, APL-2. The trials ... dose studies designed to assess the safety, tolerability, ... in healthy adult volunteers. Forty subjects ... single dose (ranging from 45 to 1,440mg) or ...
(Date:6/23/2016)... 2016 Andrew D ... Published recently in ... from touchONCOLOGY, Andrew D Zelenetz , discusses ... care is placing an increasing burden on healthcare ... therapies. With the patents on many biologics expiring, ...
Breaking Biology Technology:
(Date:6/7/2016)... 2016  Syngrafii Inc. and San Antonio Credit ... includes integrating Syngrafii,s patented LongPen™ eSignature "Wet" solution ... will result in greater convenience for SACU members ... maintaining existing document workflow and compliance requirements. ... Highlights: ...
(Date:6/1/2016)... , June 1, 2016 ... in Election Administration and Criminal Identification to Boost Global ... a recently released TechSci Research report, " Global Biometrics ... Region, Competition Forecast and Opportunities, 2011 - 2021", the ... billion by 2021, on account of growing security concerns ...
(Date:5/9/2016)... , UAE, May 9, 2016 ... it comes to expanding freedom for high net worth ... Even in today,s globally connected world, there is still ... system could ever duplicate sealing your deal with a ... second passports by taking advantage of citizenship via investment ...
Breaking Biology News(10 mins):