Navigation Links
Actinomyces viscosus

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.501 04/2002 Microorganism Actinomyces viscosus Cell type Bacteria, gram negative Molecules injected Plasmid DNA (in 1 mM Tris-HCl-0.1 mM EDTA (pH 8.0)) Growth medium Lactobacillus carrying medium (LCM) with 20 mM DL-threonine Washing solution Sterile, distilled water; sterile 10% glycerol Electroporation solution Sterile, 10% glycerol Outgrowth medium Complex medium (CAM) Cuvette 2 mm gap width Reference Yeung, M. K. and Kozelsky, C. S. 1994 Journal of Bacteriology 176. No. 13 4173-4176 Making electrocompetent cells:
  1. Grow a 500 ml cell culture in the early exponential phase until an O.D.600 of 0.2 is reached. Chill on ice for 1.5 h.
  2. 2. Harvest by centrifugation.
  3. Wash extensively first with a large volume of ice-cold sterile distilled water and then with sterile 10% glycerol.
  4. Resuspend in 10% glycerol to a final volume of 2 ml. Store in aliquots at 80 C.
Electroporation of cells:
  1. Thaw bacterial suspensions on ice. Add up to 5 l (100 ng) plasmid DNA to 100 l (5 x 108) 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) 2,500 V Time constant (T) 5 ms
  4. Dilute cells with 1 ml of CAM and incubate for 2 h at 37 C.
  5. Plate onto selective brain heart infusion agar plates; incubate at 37 C for up to 5 days.

Expected results:

Transformation efficiency up to 2.9 x 107 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Actinomyces viscosus
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/12/2017)... ... October 12, 2017 , ... The ... three Winners and six Finalists of the 2017 Blavatnik Regional Awards for Young ... Family Foundation and administered by the New York Academy of Sciences to honor ...
(Date:10/12/2017)... ... October 12, 2017 , ... ... Rosalind™, the first-ever genomics analysis platform specifically designed for life science researchers ... honor of pioneering researcher Rosalind Franklin, who made a major contribution to ...
(Date:10/11/2017)... ... October 11, 2017 , ... Proscia Inc ., ... a Webinar titled, “Pathology is going digital. Is your lab ready?” with Dr. ... best practices and how Proscia improves lab economics and realizes an increase in ...
(Date:10/11/2017)... ... October 11, 2017 , ... Singh Biotechnology today announced ... to SBT-100, its novel anti-STAT3 (Signal Transducer and Activator of Transcription 3) B ... to cross the cell membrane and bind intracellular STAT3 and inhibit its function. ...
Breaking Biology Technology:
(Date:5/23/2017)... -- Hunova, the first robotic gym for the rehabilitation and functional motor sense ... Genoa, Italy . The first 30 robots will be available ... USA . The technology was developed and patented at the IIT ... Movendo Technology thanks to a 10 million euro investment from entrepreneur Sergio ... ...
(Date:4/24/2017)... WASHINGTON , April 24, 2017 ... counsel and partner with  Identity Strategy Partners, LLP ... "With or without President Trump,s March 6, ... Foreign Terrorist Entry , refugee vetting can be instilled ... refugee resettlement. (Right now, all refugee applications are ...
(Date:4/13/2017)... MONICA, Calif. , April 13, 2017 ... New York will feature emerging and evolving ... Summits. Both Innovation Summits will run alongside the expo ... of speaker sessions, panels and demonstrations focused on trending ... coast,s largest advanced design and manufacturing event will take ...
Breaking Biology News(10 mins):