Navigation Links
Clostridium botulinum

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.509 12/2001 Microorganism Clostridium botulinum Cell type Bacteria, gram positive Molecules injected Plasmid DNA (pGK12 in water) Growth medium TPGY medium Washing solution 10% PEG 8000 Electroporation solution 10% PEG 8000 Outgrowth medium TPGY medium with 25 mM MgCl2 Cuvette 4 mm gap width Reference Zhou Y. and Johnson E. A. 1993 Biotechnology Letters 15, No. 2 121-126 Making electrocompetent cells:

1. Grow cells in TPGY medium at 37 C to an O.D.660 of 0.8. 2. Harvest by centrifugation. Wash in 0.8 volumes of PEG at 4 C, keep cells cold during the entire procedure. 3. Resuspend cells in 0.8 ml 10% PEG (number of cells: 4 x 108 cells/ml), keep on ice.

Electroporation of cells:

  1. Add 1 g plasmid DNA to 800 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into a prechilled cuvette. Keep on ice for 2 min.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,500 V Time constant (T) 5 ms
  4. Dilute cells into 10 ml TPGY medium with 25 mM MgCl2 and incubate 5 hours at 34 C.
  5. Plate cells on selective TPGY agar plates; incubate at 34 C for 2-3 days.
Expected Results: Transformation efficiency up to 3 x 103 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Clostridium botulinum
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/27/2016)... -- Global demand for enzymes is forecast to grow ... billion.  This market includes enzymes used in industrial ... animal feed, and other markets) and specialty applications ... beverages will remain the largest market for enzymes, ... containing enzymes in developing regions.  These and other ...
(Date:6/27/2016)... ... 2016 , ... Parallel 6 , the leading software as a service ... Virtual Patient Encounter CONSULT module which enables both audio and video telemedicine communication ... , Using the CONSULT module, patients and physicians can schedule a face to face ...
(Date:6/27/2016)...   Ginkgo Bioworks , a leading organism ... today awarded as one of the World Economic ... most innovative companies. Ginkgo Bioworks is engineering biology ... world in the nutrition, health and consumer goods ... customers including Fortune 500 companies to design microbes ...
(Date:6/24/2016)... ... 24, 2016 , ... Researchers at the Universita Politecnica delle Marche in Ancona ... or pleural mesothelioma. Their findings are the subject of a new article on the ... are signposts in the blood, lung fluid or tissue of mesothelioma patients that can ...
Breaking Biology Technology:
(Date:6/20/2016)... -- Securus Technologies, a leading provider of civil ... investigation, corrections and monitoring announced that after exhaustive ... the final acceptance by all three (3) Department ... (MAS) installed. Furthermore, Securus will have contracts for ... October, 2016. MAS distinguishes between legitimate wireless device ...
(Date:6/9/2016)... Paris Police Prefecture ... solution to ensure the safety of people and operations in ... major tournament Teleste, an international technology group specialised ... today that its video security solution will be utilised by ... public safety across the country. The system roll-out is scheduled ...
(Date:6/2/2016)... NEW YORK , June 2, 2016   The ... (Weather), is announcing Watson Ads, an industry-first capability in which ... advertising, by being able to ask questions via voice or ... Marketers have long ... with the consumer, that can be personal, relevant and valuable; ...
Breaking Biology News(10 mins):