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


(Date:2/11/2016)... (PRWEB) , ... February 11, 2016 , ... ... for more than 150 years, continues today to pursue the highest level of ... of analytical instruments: the AR9 Refractometer and the AR5 Refractometer. Accurate, reliable ...
(Date:2/10/2016)... Md. , Feb. 10, 2016  The Maryland ... Busch , has announced that University of Maryland School ... PhD, MBA and University of Maryland Medical System President ... of the "Speaker,s Medallion," the highest honor given to ... of Delegates. Dean Reece and Mr. ...
(Date:2/10/2016)...  Allergan plc (NYSE: AGN ) a leading ... , Allergan,s CEO and President, will be featured as ... the RBC Capital Markets Healthcare Conference on Tuesday, February ... York Palace Hotel in New York, NY ... can be accessed on Allergan,s Investor Relations web site ...
(Date:2/10/2016)... , ... February 10, 2016 , ... ... to their comprehensive training and support program, Sonalinkā„¢ remote monitoring. The inaugural launch ... performed on Friday, February 5th, connecting Dr. Samuel Peretsman to a HIFU technical ...
Breaking Biology Technology:
(Date:2/3/2016)... http://www.researchandmarkets.com/research/d8zjcd/emotion_detection ) ... "Emotion Detection and Recognition Market by Technology ... Tools (Facial Expression, Voice Recognition and Others), ... Global forecast to 2020" report to ... ) has announced the addition of the ...
(Date:2/2/2016)... YORK , Feb. 2, 2016 ... of the bioinformatic market by reviewing the recent ... enabled tools that drive the field forward. Includes ... to: Identify the challenges and opportunities that ... providers and software solution developers, as well as ...
(Date:2/2/2016)... YORK , Feb. 2, 2016 Technology Enhancements ... presents an analysis of the digital and computed radiography ... Malaysia , and Indonesia ... trends and market size, as well as regional market ... country and discusses market penetration and market attractiveness, both ...
Breaking Biology News(10 mins):