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


(Date:5/2/2016)... ... May 02, 2016 , ... F.E.E.D. Co., the ... of their revolutionary, veterinarian-designed product for indoor cats. The NoBowl Feeding System replaces ... their food the way nature intended. NoBowls make cats happy and healthy. , ...
(Date:4/29/2016)... ... April 29, 2016 , ... ... to transform technology into a viable company, CereScan’s CEO, John Kelley, joined other ... a recognized leader and mentor in the Denver area business community, shared his ...
(Date:4/28/2016)... , April 28, 2016 ... company reports the Company,s CEO  was featured in ... Accelerators Enter When VCs Fear To Tread: ... Leader magazine is an essential business ... everything from emerging biotechs to Big Pharmas. Their ...
(Date:4/27/2016)... Boston (PRWEB) , ... April 27, 2016 , ... ... driven by semantic web technology, today announced that it has been named to The ... life sciences, financial services and other markets, Cambridge Semantics serves the needs of end ...
Breaking Biology Technology:
(Date:4/28/2016)... and BANGALORE, India , April ... EdgeVerve Systems, a product subsidiary of Infosys (NYSE: ... today announced a global partnership that will provide ... to use mobile banking and payment services.      ... a key innovation area for financial services, but it also ...
(Date:4/26/2016)... 27, 2016 Research and ... Biometrics Market 2016-2020"  report to their offering.  , ... The analysts forecast the global multimodal biometrics ... during the period 2016-2020.  Multimodal biometrics ... such as the healthcare, BFSI, transportation, automotive, and ...
(Date:4/15/2016)... , April 15, 2016 ... the,  "Global Gait Biometrics Market 2016-2020,"  report to ... http://photos.prnewswire.com/prnh/20160330/349511LOGO ) , ,The global gait biometrics ... of 13.98% during the period 2016-2020. ... angles, which can be used to compute factors ...
Breaking Biology News(10 mins):