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:3/22/2017)... March 22, 2017   iSpecimen ┬«, the ... Doctors Pathology Service (DPS), a full-service anatomic ... the United States , has joined a ... Information Network (DHIN) to make human biospecimens and ... The novel program, announced in 2015 as a collaboration ...
(Date:3/22/2017)... , March 22, 2017   Boston Biomedical , ... therapeutics designed to target cancer stemness pathways, today announced ... Andrews as Chief Executive Officer, effective April 24, ... Chiang J. Li , M.D., FACP, who has led ... ago. Under his leadership, Boston Biomedical has grown from ...
(Date:3/22/2017)... ... March 21, 2017 , ... Proper glycosylation is critical for ... increase and/or decrease in antibody-dependent cellular cytotoxicity or complement-dependent cytotoxicity, there is a ... , To meet this demand, the team at SCIEX has developed a ...
(Date:3/22/2017)... ... March 21, 2017 , ... ... ) has unveiled its innovative Quantum peristaltic pump with patented ReNu single-use ... sets the new standard for high-pressure feed pumps in SU tangential flow ...
Breaking Biology Technology:
(Date:3/2/2017)... , March 2, 2017 Summary ... better understand Merck KGaA and its partnering interests and ... https://www.reportbuyer.com/product/3605601/ Description The Partnering Deals and Alliance ... partnering activity of one of the world,s leading life ... prepared upon purchase to ensure inclusion of the most ...
(Date:2/28/2017)... ANGELES , Feb. 28, 2017   Acuant ... verification software globally, announces significant enhancements to new and ... May 2016. New products include mobile and desktop Acuant ... DocX TM - a real time manual review ... core idScan┬« technology provides the fastest and most accurate ...
(Date:2/26/2017)... Feb. 25, 2017  Securus Technologies, a leading ... for public safety, investigation, corrections and monitoring, announces ... Reentry. "Too often, too many offenders ... county jails are trying to tackle this ongoing ... friends and family members. While significant steps are underway, ...
Breaking Biology News(10 mins):