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:9/25/2017)... ... 2017 , ... Cambridge Semantics , a big data ... the Big Data special edition of CIOReview magazine. , Since 2013, CIOReview ... technologies and solutions that are advancing the industry and optimizing businesses. A distinguished ...
(Date:9/25/2017)... ... September 25, 2017 , ... Having met specific requirements for ... been found to provide product development services that consistently meet customer ... clients have a lot at stake when they invest in developing ...
(Date:9/22/2017)... ... September 22, 2017 , ... The Academy of Model ... 28 and 29, 2017, to promote AMA’s programs, member services, and the model ... hobbies, including but not limited to model aviation and other RC activities. , ...
(Date:9/21/2017)... ... September 21, 2017 , ... Colpitts Clinical has launched the ... includes new features that facilitate streamlined and compliant communication, file sharing and document ... forefront of medical advancements, they rely on efficiencies to help them address the ...
Breaking Biology Technology:
(Date:4/5/2017)... , April 5, 2017  The Allen Institute for ... Cell Explorer: a one-of-a-kind portal and dynamic digital window ... imaging data, the first application of deep learning to ... stem cell lines and a growing suite of powerful ... for these and future publicly available resources created and ...
(Date:3/30/2017)... 2017  On April 6-7, 2017, Sequencing.com will host ... hackathon at Microsoft,s headquarters in Redmond, ... on developing health and wellness apps that provide a ... Genome is the first hackathon for personal genomics ... companies in the genomics, tech and health industries are ...
(Date:3/27/2017)... CENTRE, N.Y. , March 27, 2017 /PRNewswire-USNewswire/ ... Healthcare Information and Management Systems Society (HIMSS) Analytics ... Outpatient EMR Adoption Model sm . In addition, ... 12% of U.S. hospitals using an electronic medical ... CHS for its high level of EMR usage ...
Breaking Biology News(10 mins):