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.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Bifidobacterium animalis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/27/2016)... , ... June 27, 2016 , ... Parallel 6 ... trials, announced today the Clinical Reach Virtual Patient Encounter CONSULT module which ... with the physician and clinical trial team. , Using the CONSULT module, patients and ...
(Date:6/27/2016)... --  Ginkgo Bioworks , a leading organism design ... awarded as one of the World Economic Forum,s ... innovative companies. Ginkgo Bioworks is engineering biology to ... in the nutrition, health and consumer goods sectors. ... including Fortune 500 companies to design microbes for ...
(Date:6/24/2016)... NC (PRWEB) , ... June 24, 2016 , ... Researchers ... the most commonly-identified miRNAs in people with peritoneal or pleural mesothelioma. Their findings are ... to read it now. , Diagnostic biomarkers are signposts in the blood, lung ...
(Date:6/23/2016)... 23, 2016 /PRNewswire/ - FACIT has announced the ... biotechnology company, Propellon Therapeutics Inc. ("Propellon" or ... of a portfolio of first-in-class WDR5 inhibitors for ... as WDR5 represent an exciting class of therapies, ... medicine for cancer patients. Substantial advances have been ...
Breaking Biology Technology:
(Date:6/16/2016)... , June 16, 2016 ... size is expected to reach USD 1.83 billion ... Grand View Research, Inc. Technological proliferation and increasing ... applications are expected to drive the market growth. ... , The development of advanced multimodal ...
(Date:6/3/2016)... 2016 Das DOTM ... Nepal hat ein 44 Millionen ... Kennzeichen, einschließlich Personalisierung, Registrierung und IT-Infrastruktur, an ... und Implementierung von Identitätsmanagementlösungen. Zahlreiche renommierte internationale ... teilgenommen, aber Decatur wurde als konformste und ...
(Date:5/24/2016)... 24, 2016 Ampronix facilitates superior patient care by providing unparalleled technology to ... display is the latest premium product recently added to the range of products distributed ... ... ... Imaging- LCD Medical Display- Ampronix News ...
Breaking Biology News(10 mins):