Navigation Links
Saccharomyces cerevisiae

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.531 12/2001 Microorganism Saccharomyces cerevisiae Cell type Yeast Molecules injected Plasmid DNA Growth medium YEPD (1% yeast extract, 2% bactopeptone, 2% dextrose) Washing solution Ice-cold, sterile water; ice-cold 1 M sorbitol Electroporation solution Ice-cold 1 M sorbitol Outgrowth medium 1 M sorbitol Cuvette 2 mm gap width Reference Dr. Robert Sclafani University of Colorado Health Science Center Denver, CO USA Making electrocompetent cells:

1. Inoculate 500 ml YEPD with an aliquot of an overnight culture or a colony from a plate and grow to an O.D.600 of 1.3-1.5 (about 1 x 108 cells/ml). 2. Harvest by centrifugation at 4,000 x g for 5 min. at 4 C. 3. Wash in 500 ml ice-cold sterile water, centrifuge (4,000 x g, 5 min., at 4 C), repeat washing step with 250 ml water. 4. Resuspend in 20 ml ice-cold sorbitol and c entrifuge as above. Resuspend in 0.5 ml of 1 M sorbitol to a final volume of 1.0 to 1.5 ml, keep on ice.

Electroporation of cells:

  1. Add <5 l (0.1 g) DNA to 65 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Incubate 5 min on ice. 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) 1,500 V Time constant (T) 5 ms
  4. Immediately add 1 ml of cold sorbitol. Plate various aliquots onto selective plates containing 1 M sorbitol.
Expected results: Transformation efficiency up to 104 to 105 transformants/g of DNA. Note: It is possible to scale this protocol down by starting with a culture volume of 50 ml.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Saccharomyces cerevisiae
2. Saccharomyces cerevisiae
3. Saccharomyces cerevisiae
4. Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae
5. Cytosolic expression of Green Fluorescent Protein (GFP) and its derivatives in the yeast Saccharomyces cerevisiae: Detection in vivo using the Varian Cary Eclipse
6. Monitoring fluorescence resonance energy transfer (FRET) between GFP fusions in lysates of the yeast Saccharomyces cerevisiae using the Varian Cary Eclipse
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/4/2016)... China , Feb. 4, 2016 Beike ... and various medical institutions attended a ceremony in late ... integrative, personalized cell therapy in 2016. ... "Shenzhen Clinical Translation Platform for Personalized Cell Therapy" was ... Regional Cell Production Center, both subsidiaries of Beike Biotechnology ...
(Date:2/4/2016)... and MENLO PARK, Calif. , Feb. 4, ... and the "Company"), a biopharmaceutical company focused on the development ... will present at the 18 th Annual BIO ... at 10:00 a.m. EST in New York, NY ... and CEO, will provide an update on the ongoing clinical ...
(Date:2/4/2016)... Feb. 4, 2016 ContraVir Pharmaceuticals, Inc. (NASDAQ: ... development and commercialization of targeted antiviral therapies, announced today ... Conference 2016, to be held February 8-9, 2016, at ... 2016 Disruptive Growth & Healthcare Conference, taking place in ... 2016. James Sapirstein , Chief Executive Officer ...
(Date:2/4/2016)... BEIJING , Feb. 4, 2016 Sinovac ... ), a leading provider of biopharmaceutical products in ... committee of its board of directors received on February ... 3, 2016, from a consortium comprised of PKU V-Ming ... Sinobioway Biomedicine Co., Ltd., CICC Qianhai Development ( ...
Breaking Biology Technology:
(Date:2/2/2016)... Calif. , Feb. 2, 2016  Based ... market, Frost & Sullivan recognizes US-based Intelligent Retinal ... Frost & Sullivan Award for New Product Innovation. ... in North America , is ... the rapidly growing diabetic retinopathy market. The IRIS ...
(Date:1/28/2016)... Jan. 28, 2016 Synaptics (NASDAQ: SYNA ), a ... its second quarter ended December 31, 2015. ... of fiscal 2016 increased 2 percent compared to the comparable quarter ... of fiscal 2016 was $35.0 million, or $0.93 per diluted share. ... income for the first quarter of fiscal 2016 grew 9 percent ...
(Date:1/22/2016)... DUBLIN , January 22, 2016 ... has announced the addition of the  ... to their offering. --> ... of the  "Global Behavioral Biometric Market ... --> Research and Markets ( http://www.researchandmarkets.com/research/4lmf2s/global_behavioral ...
Breaking Biology News(10 mins):