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 centrifuge 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:
TAG: Saccharomyces cerevisiae

(Date:7/10/2014)... Understanding the need for better quality and higher ... that currently exists in this area, ChanTest has recently ... validated ion channel and GPCR cell lines. ChanTest’s ... channels control many critical physiological functions throughout the body. ... to treat a variety of human diseases. , ...
(Date:7/10/2014)... (PRWEB) July 10, 2014 Robert Harman, DVM, ... Medicine company, is proud to announce the relaunch of his ... with a new series called “ What are Stem Cells ... an honest and straightforward foundation in the basics of stem ... decision on the right type of treatment when considering regenerative ...
(Date:7/10/2014)... CA (PRWEB) July 10, 2014 Senior ... institutions and professional associations, addressed the challenges of “Reducing ... Medical Device Industries” through strategic sourcing. Describing the partnership ... with the Supply Chain Management Institute ( SCMI) ... event on June 24 at the Kroc Institute of ...
(Date:7/10/2014)... July 10, 2014  Kainos Capital, a firm specializing ... announced that it has acquired the Slim-Fast brand from ... business. Terms of the transaction were not disclosed. ... replacement business that markets ready-to-drink shakes, powders, bars and ... and in the United Kingdom ...
Breaking Biology Technology:Leader in Ion Channel Research Announces Newly Validated and Re-Engineered Cells 2Leader in Ion Channel Research Announces Newly Validated and Re-Engineered Cells 3Robert Harman, DVM Talks About What Stem Cells are in His Latest Blog Series for Vet-Stem, Inc. 2Robert Harman, DVM Talks About What Stem Cells are in His Latest Blog Series for Vet-Stem, Inc. 3Biotech & Biopharm Supply Chain Management Executives Launch BSMA in San Diego 2Biotech & Biopharm Supply Chain Management Executives Launch BSMA in San Diego 3Biotech & Biopharm Supply Chain Management Executives Launch BSMA in San Diego 4Kainos Capital Acquires Slim-Fast From Unilever 2
... , BEIJING, China , July 6 ... the development, assembly,marketing and sale of medical devices and homecare medical products in ... EPS for,fiscal year 2010. , , ... (Logo: http://photos.prnewswire.com/prnh/20100422/CNTH001LOGO ) , ...
... Cellectis (Alternext: ALCLS), the French genome engineering specialist, announced today ... by Taconic Artemis and Taconic Farms Inc in December 2008 ... , , , ... of the Tribunal de Grande Instance decided on Tuesday, June 29 ...
... July 6 Novavax, Inc. (Nasdaq: NVAX ... M.D., as the company,s Chief Scientific Officer who will report ... Officer of Novavax.  Previously, Dr. Glenn was Chief Scientific Officer ... international health at Johns Hopkins University,s School of Public ...
Cached Biology Technology:Dehaier Medical Systems Reaffirms Earnings Estimate of $0.80 EPS for FY 2010 2Dehaier Medical Systems Reaffirms Earnings Estimate of $0.80 EPS for FY 2010 3Dehaier Medical Systems Reaffirms Earnings Estimate of $0.80 EPS for FY 2010 4Taconic's Litigation Contesting License Termination By Cellectis Is Dismissed 2Taconic's Litigation Contesting License Termination By Cellectis Is Dismissed 3Novavax Announces Appointment of Dr. Gregory Glenn as Chief Scientific Officer 2Novavax Announces Appointment of Dr. Gregory Glenn as Chief Scientific Officer 3Novavax Announces Appointment of Dr. Gregory Glenn as Chief Scientific Officer 4
(Date:7/10/2014)... -- NXT-ID, Inc., (OTCQB: NXTD) a biometric authentication company focused on the ... was interviewed on July 1 st on the ... . Mr. Pereira discusses the company,s next generation smart wallet ... the Wocket™ aims to replace a traditional wallet with a ... tells Gino he has never heard of the Wocket ...
(Date:7/10/2014)... , July 3, 2014 Research and ... of the "Global Gesture Recognition & Touch-Less ... Forecasts to 2020" report to their offering ... Global Gesture Recognition & Touch-Less Sensing Market to ... for a while, but the companies were unable ...
(Date:7/10/2014)... July 7, 2014 According to ... "Electronic Access Control Systems Market Global Forecast, Market Share, ... global Electronic Access Control systems market was valued at ... grow at a CAGR of 12.6% from 2014 to ... million in 2019. Browse the full Electronic ...
Breaking Biology News(10 mins):Security and Your Money: Tampa Bay Radio Host Cedric Harris Talks to NXT-ID CEO Mr. Gino Pereira about the Wocket Smart Wallet 2Security and Your Money: Tampa Bay Radio Host Cedric Harris Talks to NXT-ID CEO Mr. Gino Pereira about the Wocket Smart Wallet 3Security and Your Money: Tampa Bay Radio Host Cedric Harris Talks to NXT-ID CEO Mr. Gino Pereira about the Wocket Smart Wallet 4Global Gesture Recognition & Touch-Less Sensing (2D, 3D, Ultrasonic, IR, Capacitive) Market - Forecasts to 2020 2Electronic Access Control Systems Market is Expected to Reach USD 31.2 Billion Globally in 2019: Transparency Market Research 2Electronic Access Control Systems Market is Expected to Reach USD 31.2 Billion Globally in 2019: Transparency Market Research 3Electronic Access Control Systems Market is Expected to Reach USD 31.2 Billion Globally in 2019: Transparency Market Research 4Electronic Access Control Systems Market is Expected to Reach USD 31.2 Billion Globally in 2019: Transparency Market Research 5
... Entering a new domain . . . ... at Oak Ridge National Laboratory,s Center for Nanophase ... discoveries that could open new pathways for nanoscale ... ( http://www.nature.com/nmat/journal/vaop/ncurrent/abs/nmat2373 ) and led by Ramamoorthy ...
... physical and emotional toll on our bodies and scientists are ... we respond to stress at the cellular level in the ... stress is essential for survival. Using a rat ... his team of researchers at the Hotchkiss Brain Institute have ...
... across the country will convene on the University of Houston ... public health issues and intervention strategies. , The ... April 9 10 at the UH M.D. Anderson Library, ... inaugural conference and have high hopes that others will see ...
Cached Biology News:Story tips from the Departments of Energy's Oak Ridge National Lab -- March 2009 2Story tips from the Departments of Energy's Oak Ridge National Lab -- March 2009 3New and unexpected mechanism identified how the brain responds to stress 2Texas Obesity Research Center at UH assembles researchers to discuss obesity 2
Monocarboxylate (lactate) transporter 2...
Mouse Anti-Human Kallikrein 2 (Hk2)...
... (Ankyrin repeat and SOCS box-containing protein) This ... containing amino acids 75-93 and 348-368 of ... Catalog No. 40149) can be used as ... with mouse and rat Asb-2, based on ...
Recognizes TWIK-2. Does not cross react with any other known proteins....
Biology Products: