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:4/27/2017)... ... April 27, 2017 , ... The ... Communication Award goes to Jayson Lusk, a consummate communicator who promotes agricultural science ... advocate for science, as he explains how innovation and growth in agriculture are ...
(Date:4/26/2017)... (PRWEB) , ... April 26, 2017 , ... ... FDA’s GMP expectations for phase I clinical trials comes to Tampa, San Francisco ... biotechnology and pharma professionals representing FDA regulated organizations such as Pfizer Inc., Teva ...
(Date:4/26/2017)... HATFIELD, Pa. , April 26, 2017 /PRNewswire/ ... drug delivery platform, has signed a collaborative and ... through Dr. Silvia Muro . The overall ... pharmacokinetics and pharmacodynamics of various 3DNA designs and ... aims involve targeting diseases of the vasculature as ...
(Date:4/25/2017)... , ... April 25, 2017 , ... ... Physical Medicine & Rehabilitation, P.A. , proudly announced today that acclaimed physiatrist Matthew ... his duties on May 15, 2017. , Dr. Terzella completed his residency in ...
Breaking Biology Technology:
(Date:3/9/2017)... and MOUNTAIN VIEW, Calif. , March ... Made Simple," and 23andMe , the leading personal ... food choices.  Zipongo can now provide customers with personalized ... health goals and biometrics, but also genetic markers impacting ... Zipongo,s personalized food decision support platform uses ...
(Date:3/7/2017)...   HireVue , the leading provider of video ... best talent, faster, today announced the additions of ... Diana Kucer as Chief Marketing Officer (CMO). ... poised to drive continued growth in the company,s new ... record bookings in 2017. "Companies worldwide turn ...
(Date:3/2/2017)... -- Summary This report provides all the information ... and activities since 2010. ... Read the full report: http://www.reportlinker.com/p03605615-summary/view-report.html ... provides an in-depth insight into the partnering activity of one ... demand company reports are prepared upon purchase to ensure inclusion ...
Breaking Biology News(10 mins):