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:10/12/2017)... Irvine, ca (PRWEB) , ... October 12, 2017 ... ... for the Surgical Wound Market with the addition of its newest module, US ... the $1.2B market for thrombin hemostats, absorbable hemostats, fibrin sealants, synthetic sealants and ...
(Date:10/11/2017)... ... ... a basic first aid supply for any work environment, but most personal eye wash can ... a dangerous substance enters both eyes? It’s one less decision, and likely quicker response time ... , “Whether its dirt and debris, or an acid or alkali, getting anything in your ...
(Date:10/11/2017)... ... 11, 2017 , ... Disappearing forests and increased emissions are the main causes ... each year. Especially those living in larger cities are affected by air pollution related ... the most pollution-affected countries globally - decided to take action. , “I knew I ...
(Date:10/10/2017)... PA (PRWEB) , ... October 10, 2017 , ... ... City Science Center’s FirstHand program has won a US2020 STEM Mentoring Award. Representatives ... the award for Excellence in Volunteer Experience from US2020. , US2020’s mission is ...
Breaking Biology Technology:
(Date:3/28/2017)... , March 28, 2017 ... Biometrics), Hardware (Camera, Monitors, Servers, Storage Devices), Software (Video ... and Region - Global Forecast to 2022", published by ... in 2016 and is projected to reach USD 75.64 ... 2017 and 2022. The base year considered for the ...
(Date:3/24/2017)... The Controller General of Immigration from Maldives Mr. Mohamed Anwar ... the prestigious international IAIR Award for the most innovative high security ePassport ... ... Maldives Immigration Controller General, Mr. ... on the right) have received the IAIR award for the "Most innovative ...
(Date:3/23/2017)... The report "Gesture Recognition and Touchless Sensing Market by Technology (Touch-based and ... 2022", published by MarketsandMarkets, the market is expected to be worth USD 18.98 ... Continue Reading ... ...      ...
Breaking Biology News(10 mins):