Navigation Links
Propionibacterium freudenreichii

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.527 04/2002 Microorganism Propionibacterium freudenreichii Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium SLB medium Washing solution Ice-cold 0.5 M sucrose Electroporation solution Ice-cold 0.5 M sucrose buffered with 1 mM potassium acetate (pH 5.5) Outgrowth medium SLB medium with 0.5 M sucrose Cuvette 1 mm gap width Reference Jore, J. P M. et al 2001 Applied and Environmental Microbiology 67, No. 2 499-503 Making electrocompetent cells:

1. Cultivate cells anaerobically at 30 C in SLB medium to the stationary growth phase. Dilute 1:50 in fresh SLB medium and incubate again for about 20 h. 2. Harvest in the exponential growth phase by centrifugation and wash extensively with ice-cold 0.5 M sucrose. 3. Wash once in ice-cold electroporation solution. 4. Resuspend in ice-cold electroporation solution (abo ut 1/100 of the original culture volume).

Electroporation of cells:

  1. Add plasmid DNA to 80-100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,000 V Time constant (T) 5 ms
  4. Immediately add 900 l cold outgrowth medium and incubate for 2.5-3 h at 30 C.
  5. Plate cells onto selective SLB plates; incubate 5-7 days at 30 C under anaerobic conditions.
Expected Results: Transformation efficiency up to 1 x 108 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Propionibacterium freudenreichii
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... , June 23, 2016 A person commits ... the crime scene to track the criminal down. ... U.S. Food and Drug Administration (FDA) uses DNA evidence to ... Sound far-fetched? It,s not. The FDA has ... to support investigations of foodborne illnesses. Put as simply as ...
(Date:6/23/2016)... FRANCISCO , June 23, 2016   EpiBiome ... has secured $1 million in debt financing from Silicon ... ramp up automation and to advance its drug development ... its new facility. "SVB has been an ... beyond the services a traditional bank would provide," said ...
(Date:6/23/2016)... Calif. , June 23, 2016  Blueprint Bio, ... biological discoveries to the medical community, has closed its ... Matthew Nunez . "We have received ... with the capital we need to meet our current ... essentially provide us the runway to complete validation on ...
(Date:6/23/2016)... ... June 23, 2016 , ... ... line of intelligent tools designed, tuned and optimized exclusively for Okuma CNC machining ... Chicago. The result of a collaboration among several companies with expertise in toolholding, ...
Breaking Biology Technology:
(Date:6/21/2016)... VANCOUVER, British Columbia , June 21, 2016 ... been appointed to the new role of principal ... has been named the director of customer development. ... , NuData,s chief technical officer. The moves reflect ... development teams in response to high customer demand ...
(Date:6/16/2016)... 2016 The global ... reach USD 1.83 billion by 2024, according to ... Technological proliferation and increasing demand in commercial buildings, ... drive the market growth.      (Logo: ... development of advanced multimodal techniques for biometric authentication ...
(Date:6/9/2016)... control systems is proud to announce the introduction of fingerprint attendance control software, allowing ... are actually signing in, and to even control the opening of doors. ... ... ... Photo - http://photos.prnewswire.com/prnh/20160609/377487 ...
Breaking Biology News(10 mins):