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:6/27/2016)... ... 27, 2016 , ... Cancer experts from Austria, Hungary, Switzerland, ... a new and helpful biomarker for malignant pleural mesothelioma. Surviving Mesothelioma has just ... now. , Biomarkers are components in the blood, tissue or body fluids ...
(Date:6/27/2016)... , June 27, 2016  Liquid Biotech ... announced the funding of a Sponsored Research Agreement ... circulating tumor cells (CTCs) from cancer patients.  The ... in CTC levels correlate with clinical outcomes in ... These data will then be employed to support ...
(Date:6/24/2016)... NY (PRWEB) , ... June 24, 2016 , ... While ... machines such as the Cary 5000 and the 6000i models are higher end machines ... is the height of the spectrophotometer’s light beam from the bottom of the cuvette ...
(Date:6/23/2016)... Mass. , June 23, 2016   ... development of novel compounds designed to target cancer ... napabucasin, has been granted Orphan Drug Designation from ... the treatment of gastric cancer, including gastroesophageal junction ... stemness inhibitor designed to inhibit cancer stemness pathways ...
Breaking Biology Technology:
(Date:3/23/2016)... WAKEFIELD, Massachusetts , March 23, 2016 ... kombiniert im Interesse erhöhter Sicherheit Gesichts- und ... Xura, Inc. (NASDAQ: MESG ... heute bekannt, dass das Unternehmen mit SpeechPro ... insbesondere aus der Finanzdienstleistungsbranche, wird die Möglichkeit ...
(Date:3/22/2016)... PROVO and SANDY, Utah ... Ontario (NSO), which operates the highest sample volume laboratory ... and Tute Genomics and UNIConnect, leaders in clinical sequencing ... announced the launch of a project to establish the ... panel. NSO has been contracted by ...
(Date:3/18/2016)... LONDON , March 18, 2016 ... Established Suppliers of Biometrics, ICT, Manned & Unmanned Vehicles, Physical ... & security companies in the border security market and ... and Europe has led ... your companies improved success. --> defence & ...
Breaking Biology News(10 mins):