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:4/27/2017)... NASHVILLE, Tenn. , April 27, 2017  Pendant ... company developing innovative surface modification and drug delivery technologies, ... & Johnson Innovation, JLABS @ Toronto ... Chief Executive Officer of Pendant Biosciences, noted, "We are ... Toronto community, and are honored to ...
(Date:4/26/2017)... ... ... LABS, Inc. (LABS) announced in December 2016 that two new Zika ... Testing (NAT) for ZIKV; and Enzyme Immunoassays (EIAs) specific for IgM and IgG ZIKV ... donors under an Investigational New Drug (IND) study protocol. , Now, as part ...
(Date:4/25/2017)... ... April 25, 2017 , ... As part of the Stago ... the laboratory testing for DIC in order to illuminate this clinical problem for people ... in hospitalized patients resulting in a high degree of morbidity and mortality. DIC is ...
(Date:4/25/2017)... ... April 25, 2017 , ... Covalent Metrology ... Analytical Services unit provides high-quality data to clients, both faster and cheaper ... receipt. There are no price premiums, and customers are welcome to participate in ...
Breaking Biology Technology:
(Date:4/6/2017)... Forecasts by Product Type (EAC), ... End-Use (Transportation & Logistics, Government & Public Sector, Utilities ... Generation Facility, Nuclear Power), Industrial, Retail, Business Organisation (BFSI), ... you looking for a definitive report on the $27.9bn ... ...
(Date:4/5/2017)... KEY FINDINGS The global market ... CAGR of 25.76% during the forecast period of 2017-2025. ... for the growth of the stem cell market. ... MARKET INSIGHTS The global stem cell market is segmented ... The stem cell market of the product is segmented ...
(Date:3/30/2017)... LOS ANGELES , March 30, 2017  On ... Hack the Genome hackathon at ... This exciting two-day competition will focus on developing health ... experience. Hack the Genome is ... has been tremendous. The world,s largest companies in the ...
Breaking Biology News(10 mins):