Navigation Links
Kluyveromyces lactis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.516 12/2001 Microorganism Kluyveromyces lactis AWJ137 Cell type Yeast Molecules injected DNA Growth medium Not given Washing solution YPD, 10 mM DTT, 40 mM HEPES (pH 8.0) Electroporation solution 2.5 mM Tris/HCl, 1 mM MgCl2, (pH 8.0) Outgrowth medium YPD with 1 M sorbitol Cuvette 4 mm gap width Reference Prof. Friedhelm Meinhardt Universitt Mnster Corrensstr. 3 D-48149 Mnster
e-mail: meinhar@uni-muenster.de Making electrocompetent cells:

1. Grow a 50 ml cell culture of Kluyveromyces lactis to a density of 5 x 107 cells/ml. 2. Harvest by centrifugation (5 min., at 4,000 x g) 3. Resuspend cells in 5 ml washing solution and incubate for 5 min. 4. Wash twice with electroporation solution and resuspend in 0.25 ml of the same solution. Incubate for 1 hour on ice.

Electroporation of cells:

  1. Add 5 l DNA (1 g/l) to 100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into a 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. Add 1 ml YPD medium with 1 M sorbitol, incubate for 1 hour at 30 C.
  5. Centrifuge cells 30 sec., resuspend in 0.4 ml sterile water, plate in aliquots on selective plates.
Expected results: Transformation efficiency up to 200 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

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


(Date:6/23/2016)... 23, 2016 Houston Methodist Willowbrook Hospital ... Sports Association to serve as their official health ... Methodist Willowbrook will provide sponsorship support, athletic training ... association coaches, volunteers, athletes and families. ... Sports Association and to bring Houston Methodist quality ...
(Date:6/23/2016)... YORK , June, 23, 2016  The Biodesign ... to envision new ways to harness living systems and ... Modern Art (MoMA) in New York City ... than 130 participating students, showcased projects at MoMA,s Celeste ... Paola Antonelli , MoMA,s senior curator of architecture ...
(Date:6/23/2016)... ... 2016 , ... STACS DNA Inc., the sample tracking software company, today announced ... has joined STACS DNA as a Field Application Specialist. , “I am thrilled ... COO of STACS DNA. “In further expanding our capacity as a scientific integrator, Hays ...
(Date:6/23/2016)... 2016  Blueprint Bio, a company dedicated to identifying, ... community, has closed its Series A funding round, according ... "We have received a commitment from Forentis Fund ... to meet our current goals," stated Matthew Nunez ... to complete validation on the current projects in our ...
Breaking Biology Technology:
(Date:5/20/2016)... May 20, 2016  VoiceIt is excited to ... VoicePass. By working together, VoiceIt and ... VoiceIt and VoicePass take slightly different approaches to ... both security and usability. ... this new partnership. "This marketing and ...
(Date:5/3/2016)... 3, 2016  Neurotechnology, a provider of high-precision ... Automated Biometric Identification System (ABIS) , a complete ... MegaMatcher ABIS can process multiple complex biometric transactions ... of fingerprint, face or iris biometrics. It leverages ... and MegaMatcher Accelerator , which have been ...
(Date:4/26/2016)... -- Research and Markets has announced the ...  report to their offering.  , ,     (Logo: ... forecast the global multimodal biometrics market to grow ... 2016-2020.  Multimodal biometrics is being implemented ... healthcare, BFSI, transportation, automotive, and government for controlling ...
Breaking Biology News(10 mins):