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:10/11/2017)... ... October 11, 2017 , ... Personal eye wash is a basic first aid supply for any ... So which eye do you rinse first if a dangerous substance enters both eyes? It’s ... Wash with its unique dual eye piece. , “Whether its dirt and debris, or ...
(Date:10/11/2017)... ... 2017 , ... Disappearing forests and increased emissions are the main causes of ... year. Especially those living in larger cities are affected by air pollution related diseases. ... most pollution-affected countries globally - decided to take action. , “I knew I had ...
(Date:10/10/2017)... ... October 10, 2017 , ... For the second ... a US2020 STEM Mentoring Award. Representatives of the FirstHand program travelled to Washington, ... from US2020. , US2020’s mission is to change the trajectory of STEM education ...
(Date:10/10/2017)... ... October 10, 2017 , ... ... 13 prestigious awards honoring scientists who have made outstanding contributions to ... symposium during Pittcon 2018, the world’s leading conference and exposition for laboratory science, ...
Breaking Biology Technology:
(Date:7/20/2017)... 20, 2017 Delta (NYSE: DAL ) customers now ... aircraft at Reagan Washington National Airport (DCA). ... Delta launches biometrics to board aircraft at ... Delta,s biometric boarding pass experience ... is now integrated into the boarding process to allow eligible Delta SkyMiles ...
(Date:6/14/2017)...  IBM (NYSE: IBM ) is introducing several innovative ... to developing collaboration between startups and global businesses, taking place ... the event, nine startups will showcase the solutions they have ... France is one ... a 30 percent increase in the number of startups created ...
(Date:5/16/2017)... , May 16, 2017   Bridge ... health organizations, and MD EMR Systems , ... development partner for GE, have established a partnership ... Portal product and the GE Centricity™ products, including ... EMR. These new integrations will ...
Breaking Biology News(10 mins):