Navigation Links
Lactococcus lactis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.519 12/2001 Microorganism Lactococcus lactis MG1363 Cell type Bacteria, gram positive Molecules injected Plasmid DNA (pGK12) Growth medium Complex medium with 1% glycine Washing solution 0.5 M sucrose, 10% glycerol Electroporation solution 0.5 M sucrose, 10% glycerol Outgrowth medium Ice-cold complex medium with 0.5 M sucrose, 20 mM MgCl2, 2 mM CaCl2 Cuvette 1 mm gap width Reference Dr. Horst Neve Bundesanstalt fr Milchforschung Institut fr Mikrobiologie Hermann-Weigmann Str. 1 D-24103 Kiel Phone +49 431 6091 Fax +49 431 609222 Making electrocompetent cells:

1. Grow cells overnight at 30 C to an O.D.620 of 0.7. 2. Wash twice with ice-cold washing solution. 3. Resuspend cells in 1/100 volume of electroporation solution. Keep on ice.

Electroporation of cells:

  1. Add 0.25 g plasmid DNA (in water) to 100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into 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. Add 1 ml ice-cold complex medium, incubate 2 hours at 30 C.
  5. Plate diluted cells on selective chloramphenicol plates. Incubate 2 days at 30 C.
Expected Results: Transformation efficiency up to 1.4 x 106 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Lactococcus lactis
2. Kluyveromyces lactis
3. Kluyveromyces lactis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/10/2016)... Feb. 10, 2016  Matchbook, Inc., a company ... growing biotech companies, announced today the appointment of ... Advisor. Jim brings nearly 25 years of experience ... having spent nearly two decades in executive level ... Procurement at Genzyme and, most recently headed global ...
(Date:2/10/2016)... ... February 10, 2016 , ... HOLLOWAY AMERICA, a leading custom ... Rocky Mountain Chapter 21st Annual Vendor Exhibition on Thursday, February 18, 2016. The ... for its annual event, which will run from 3:00 p.m. - 8:30 p.m. ...
(Date:2/10/2016)... ... February 10, 2016 , ... ... comprehensive training and support program, Sonalink™ remote monitoring. The inaugural launch of this ... Friday, February 5th, connecting Dr. Samuel Peretsman to a HIFU technical expert at ...
(Date:2/10/2016)... ... 10, 2016 , ... Curoverse announced today that the open-source, ... capabilities for managing and processing genomic and health data at petabyte scale. ... and analyzing genomic data,” said Adam Berrey chief executive officer at Curoverse. “Microsoft ...
Breaking Biology Technology:
(Date:2/3/2016)... , Feb. 3, 2016 ... addition of the "Emotion Detection and ... Learning, and Others), Software Tools (Facial Expression, ... End Users,and Regions - Global forecast to ... --> http://www.researchandmarkets.com/research/d8zjcd/emotion_detection ) has announced ...
(Date:2/2/2016)... , Feb. 2, 2016 Technology Enhancements Accelerate Growth ... analysis of the digital and computed radiography markets in ... , and Indonesia (TIM). It ... market size, as well as regional market drivers and ... discusses market penetration and market attractiveness, both for digital ...
(Date:2/1/2016)... 1, 2016  Today, the first day of American ... to develop a first of its kind workplace health ... Watson. In the first application of Watson ... IBM ), and Welltok will create a new offering ... cognitive analytics, delivered on Welltok,s health optimization platform. The ...
Breaking Biology News(10 mins):