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:3/23/2017)... , March 23, 2017  Northwest Biotherapeutics (OTCQB: ... personalized immune therapies for solid tumor cancers, today ... million financing it announced last Friday, March 17, ... several institutional investors securities totaling 28,843,692 shares, comprised ... and 10,000,000 shares of Class C Warrants pre-funded ...
(Date:3/23/2017)... 2017 Kineta, Inc., a biotechnology company ... in immuno-oncology, today announced the discovery and characterization ... that activate interferon response factor 3 (IRF3) via ... tumor regression in a murine colon carcinoma mouse ... complete tumor regression to initial drug treatment were ...
(Date:3/23/2017)... New Orleans, La. (PRWEB) , ... March 23, ... ... of real-time, industrial monitoring solutions, today announced the hire of Dr. Sigmund “Sig” ... for APMT customer applications, strategic partnerships and joint development activities. , “Dr. Floyd’s ...
(Date:3/22/2017)... Good Start Genetics, a leading family genomics ... million covered lives mark through its most recent payor ... . With newly signed contracts nationally and others ... payor acceptance based on the quality of its science, ... its industry-leading customer care and support and its published ...
Breaking Biology Technology:
(Date:3/23/2017)... Research and Markets has announced the addition of the "Global ... 2025" report to their offering. ... The Global Vehicle Anti-Theft System Market is ... next decade to reach approximately $14.21 billion by 2025. ... all the given segments on global as well as regional levels ...
(Date:3/20/2017)... At this year,s CeBIT Chancellor Dr. Angela ... The Chancellor came to the DERMALOG stand together with the Japanese Prime ... partner country. At the largest German biometrics company the two government leaders ... iris recognition as well as DERMALOG´s multi-biometrics system.   ... ...
(Date:3/9/2017)... , Australia , March 9, ... study data at the prestigious World Lung Imaging Workshop ... Andreas Fouras , was invited to deliver the ... pulmonary medicine. This globally recognised event brings together leaders ... share the latest developments in lung imaging. ...
Breaking Biology News(10 mins):