Navigation Links
Bifidobacterium animalis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.538 04/2002 Microorganism Bifidobacterium animalis Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium MRS broth with 0.05% cysteine.HCl and 0.5 M sucrose (final concentrations) Washing solution 0.5 M sucrose Electroporation solution 0.5 M sucrose, 1 mM ammonium citrate, pH 6 Outgrowth medium MRS broth with 0.05% cysteine.HCl and 0.5 M sucrose (final concentrations) Cuvette 1 mm gap width Reference Argnani, A. et al 1996 Microbiology 142 109-114 Making electrocompetent cells:

1. Cultivate cells by using an overnight culture to inoculate fresh medium. Grow cells overnight at 37 C. Dilute this culture 1:25 in fresh medium and cultivate at 37 C until an O.D.695 of 0.2. Chill bacteria on ice. 2. Harvest by centrifugation. 3. Wash twice with 0.5 M sucrose 4. Resuspend in about 1/250 of the original culture volume of 1 mM ice-cold sucrose-citrate buffer, dispense in tubes and incubate for 3.5 hours at 4 C.

Electroporation of cells:

  1. Add 0.5-1.5 g plasmid DNA to 80 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) 1,200 V Time constant (T) 5 ms
  4. Dilute with 800 l outgrowth medium and incubate for 2.5 h at 37 C.
  5. Plate onto selective MRS agar plates; incubate anaerobically for 2-3 days at 37 C.
Expected Results: Transformation efficiency up to 9.4 x 104 transformants/g of DNA.



Page: All 1 2

Related biology technology :

1. Bifidobacterium animalis
Post Your Comments:

(Date:10/10/2017)... SANTA BARBARA, CALIFORNIA (PRWEB) , ... October 10, ... ... risk management, technological innovation and business process optimization firm for the life sciences ... the BoxWorks conference in San Francisco. , The presentation, “Automating GxP ...
(Date:10/9/2017)... , ... October 09, 2017 , ... At its national ... Christopher Stubbs, a professor in Harvard University’s Departments of Physics and Astronomy, has been ... a member of the winning team for the 2015 Breakthrough Prize in Fundamental physics ...
(Date:10/7/2017)... ... October 06, 2017 , ... ... ProxiMeta™ Hi-C metagenome deconvolution product, featuring the first commercially available Hi-C kit. ... to perform Hi-C metagenome deconvolution using their own facilities, supplementing the company’s ...
(Date:10/5/2017)... ... October 05, 2017 , ... LabRoots , the leading provider of ... world, is giving back to cancer research with a month-long promotion supporting the advancement ... October 31, shoppers can use promo code PinkRibbon to get 10 percent off their ...
Breaking Biology Technology:
(Date:4/4/2017)... 4, 2017   EyeLock LLC , a leader ... United States Patent and Trademark Office (USPTO) has issued ... linking of an iris image with a face image ... the company,s 45 th issued patent. ... timely given the multi-modal biometric capabilities that have recently ...
(Date:3/30/2017)... , March 30, 2017 Trends, opportunities ... (physiological and behavioral), by technology (fingerprint, AFIS, iris recognition, ... recognition, and others), by end use industry (government and ... immigration, financial and banking, and others), and by region ... , Asia Pacific , and ...
(Date:3/24/2017)... 24, 2017 The Controller General of Immigration from ... Abdulla Algeen have received the prestigious international IAIR Award for the ... Continue Reading ... ... Controller Abdulla Algeen (small picture on the right) have received the IAIR ...
Breaking Biology News(10 mins):