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:3/29/2017)... , March 29, 2017  Halozyme Therapeutics, Inc. ... and drug-delivery therapies, today announced that an Oncologic ... Drug Administration voted 11 to 0 that the ... injection was favorable for patients in the proposed ... and chronic lymphocytic leukemia. The FDA action date ...
(Date:3/29/2017)... (PRWEB) , ... March 29, 2017 , ... Bactana Animal ... hormones in the world’s food supply through enhancement of the gut microbiota, today announced ... included Connecticut Innovations , New York-based Sustainable Income Capital Management, LLC and a ...
(Date:3/29/2017)... 2017 /PRNewswire/ -  GeneNews Limited (TSX:GEN) ("GeneNews" or the ... new risk stratification test for breast cancer, via its ... ("IDL"). BreastSentry incorporates a blood-based biomarker test with a sophisticated ... developing breast cancer.   ... BreastSentry measures the fasting plasma levels of ...
(Date:3/28/2017)... ... March 28, 2017 , ... NetDimensions announced today that ... enhance training plan management for consistent implementation of standards and regulatory requirements across ... Group to help improve and streamline their training and employee development programs, which ...
Breaking Biology Technology:
(Date:3/7/2017)...   HireVue , the leading provider of video ... best talent, faster, today announced the additions of ... Diana Kucer as Chief Marketing Officer (CMO). ... poised to drive continued growth in the company,s new ... record bookings in 2017. "Companies worldwide turn ...
(Date:3/2/2017)... , March 2, 2017 Who risk ... lawsuits? Download the full report: https://www.reportbuyer.com/product/4313699/ ... THE FINGERPRINT SENSOR FIELD? Fingerprint sensors using capacitive ... The fingerprint sensor vendor Idex forecasts an increase of ... mobile devices and of the fingerprint sensor market between ...
(Date:2/28/2017)... News solutions for biometrics, bag drop and New ADA-compliant kiosk ... At PTE 2017 ... Materna will present its complete end-to-end passenger journey, from ... benefit for passengers. To accelerate the whole passenger handling process, ... to take passengers through the complete integrated process with a ...
Breaking Biology News(10 mins):