Navigation Links
Using the VersaFluor Fluorometer to Quantitate GUS Expression in Plant Tissues Bombarded With the Biolistic PDS-1000/He System

plasmid (Ow et al., 1986; see Koehler et al., 1996, for details of promoter construction). The GUS gene is fused to an enhanced 35S promoter (Kay et al., 1987). An accurate measure of the concentration of the gene constructs is determined with the VersaFluor fluorometer using the Hoechst 33258 dye from Bio-Rad. The two constructs are then co-precipitated at constant and equal-molar concentrations onto gold particles (Figure 1A). Plant material is bombarded with the gold particles using the Biolistic PDS-1000/He particle gun (Bio-Rad) as depicted in Figure 1B.

Following particle bombardment and a 48-hour incubation period in ethylene (a plant hormone that induces abscission), approximately 1 mm of tissue is harvested from the upper surface of the bombarded explants (abscission zones, petioles and stems), and frozen separately in liquid nitrogen (Figure 1B). The protein extraction procedure is modified from the original protocol described by Jefferson et al. (1987) to allow the assay of cellulase, luciferase, and GUS activity from the same tissue extract. Approximately 150 mg of plant tissue is ground in liquid nitrogen, thawed in 450 l of extraction buffer (0.1 M sodium phosphate, pH 7.8, 1% Triton X-100, 2 mM EDTA, 1 mM DTT and 0.1% BSA), vortexed and then centrifuged at 10,000 rpm in a microcentrifuge for 4 minutes.

The supernatant is then transferred to new tubes in 75 l aliquots for measurement of cellulase and luciferase activity as described elsewhere (Koehler et al., 1996). For GUS assay, 75 l of the supernatant is combined with 25 l of a GUS adjustment buffer (0.4 M citrate buffer pH 5.5, 34 mM EDTA and 17 mM DTT) and the combined mixtures incubated at 55 C for 3
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
2. Oil-Free PCR Using the Hot Top Assembly
3. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
4. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
5. Using XL1-Red Mutator Strain to Generate Esterase Variants
6. Detect Released Oligosaccharides Using the Eagle Eye II Still Video Imaging System
7. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
8. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
9. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
10. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
11. Protocol for RNA Isolation Using TRIzol Reagent with Phase Lock Gel-Heavy
Post Your Comments:
(Date:8/26/2015)... -- The Diabetes Research Institute (DRI), a Center of ... of Medicine, announced today that the first patient ... first time a novel transplant technique for insulin-producing ... I/II study builds upon decades of progress in ... step toward the development of the DRI BioHub, ...
(Date:8/26/2015)... and BEIJING , August 26, 2015 ... Impressive Acheivements  DiaCardio wins 1 st place  ... The competition, led in Israel by venture ... startups from countries including the US, China , ... Latin America .  Impressive achievement for ...
(Date:8/26/2015)... 2015 Despite recent market ... optimism in biotech,s forecast is the recent run ... advancements.  Active biotechnology & healthcare companies with recent ... PBIO), Sequenom, Inc. (NASDAQ: SQNM ), Sarepta ... Corp. (NASDAQ: RPTP ) and EnteroMedics Inc. ...
(Date:8/25/2015)... 25, 2015   WuXi PharmaTech (Cayman) Inc. (NYSE: ... technology platform company serving the pharmaceutical, biotechnology, and medical ... Hong Kong Exchange Stock Code: 0950), a research-based biopharmaceutical ... with 20 years of operations in China ... signed an agreement whereby WuXi,s Laboratory Testing Division (LTD) ...
Breaking Biology Technology:Diabetes Research Institute Successfully Transplants First Patient In Pilot BioHub Trial 2Diabetes Research Institute Successfully Transplants First Patient In Pilot BioHub Trial 3Diabetes Research Institute Successfully Transplants First Patient In Pilot BioHub Trial 4Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 2Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 3Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 4Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 5Israeli Startup Takes First Place at Shengjing Global Innovation Awards 2015 6New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 2New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 3New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 4New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 5New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 6New Collaborations & Mergers For Research of Molecular Biomarkers in Human Diseases & Clinical Trials for Cancer Therapies Fueling Growth in Multi-Billion Dollar Life Sciences Market 7WuXi PharmaTech to Be Exclusive Supplier of Laboratory Testing Services to Lee's Pharm 2WuXi PharmaTech to Be Exclusive Supplier of Laboratory Testing Services to Lee's Pharm 3
... FAIRFIELD, N.J., July 17 Arno Therapeutics,Inc. (OTC ... ticker,symbol on the OTC Bulletin Board has changed ... About Arno Therapeutics, Arno Therapeutics, Inc. is ... products for the treatment of,cancer patients. Arno,s lead ...
... July 17 SAFC Supply Solutions(R),a focus ... Sigma-Aldrich Group (Nasdaq:,SIAL), announced today that the ... to assist its customers with their regulatory,requirement ... for chemicals --,Registration, Evaluation, Authorization and restriction ...
... July 16 Johnson & Johnson Pharmaceutical,Research & ... and Drug,Administration,s (FDA) Anti-Infective Drugs Advisory Committee voted ... DORIBAX(TM) (doripenem for,injection) for the treatment of hospital-acquired ... The committee voted that 500 mg of ...
Cached Biology Technology:Arno Therapeutics Announces New Ticker Symbol 2SAFC Supply Solutions Launches REACH-Compliant Products and Support 2SAFC Supply Solutions Launches REACH-Compliant Products and Support 3FDA Advisory Committee Provides Opinion of DORIBAX(TM) for the Treatment of Hospital-Acquired Pneumonia 2FDA Advisory Committee Provides Opinion of DORIBAX(TM) for the Treatment of Hospital-Acquired Pneumonia 3FDA Advisory Committee Provides Opinion of DORIBAX(TM) for the Treatment of Hospital-Acquired Pneumonia 4
(Date:8/26/2015)... The report "Multi-Factor Authentication (MFA) Market ... (Travel & Immigration, Government, Banking, Defense, Commercial Security, Consumer ... published by MarketsandMarkets, Multi-Factor Authentication Market is expected to ... CAGR of 17.7% between 2015 and 2020. ... spread through 169 P ages and ...
(Date:8/20/2015)... Aug. 20, 2015   Sensory, Inc ., ... the user experience and security of consumer electronics ... announced that its TrulySecure™ is the ... to be FIDO Certified™. The FIDO ... with the FIDO UAF (Universal Authentication Framework) 1.0 ...
(Date:8/17/2015)... , Aug. 17, 2015  NXT-ID, Inc. (NASDAQ: ... authentication company focused on the growing mobile commerce market ... new sales campaign for Q4 2015. The new marketing ... team consisting of Amplitude Marketing Group ( http://amplitudemarketing.com/ ) ... Group is a leader in retail driven marketing and ...
Breaking Biology News(10 mins):Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 2Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 3Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 4Multi-Factor Authentication Market Worth 9.60 Billion USD by 2020 5TrulySecure From Sensory Becomes First Face and Voice Biometrics Technology to be FIDO UAF Certified 2TrulySecure From Sensory Becomes First Face and Voice Biometrics Technology to be FIDO UAF Certified 3TrulySecure From Sensory Becomes First Face and Voice Biometrics Technology to be FIDO UAF Certified 4TrulySecure From Sensory Becomes First Face and Voice Biometrics Technology to be FIDO UAF Certified 5NXT-ID, Inc. Launches Sales Campaign for Wocket Smart Wallet with Award Winning Marketing Team 2NXT-ID, Inc. Launches Sales Campaign for Wocket Smart Wallet with Award Winning Marketing Team 3
... 10, 2008; Saranac Lake, N.Y. -- Findings just published ... at the Trudeau Institute shed new light on how ... prompts specific influenza-fighting cells to the lung airways during ... responsible for hundreds of thousands of hospitalizations and as ...
... from the University of Copenhagen and the Biocentre at the ... part of the immune system,s key codes. The same ... autumn, and have now put together a comprehensive picture of ... outside our cells. According to the researchers this new ...
... the Earth, thriving by filling every nook and cranny ... arrived on the scene. Now, this ability of ... food sources may play a significant role in bailing ... the Biodesign Institute,s Bruce Rittmann, Rosa Krajmalnik-Brown, and Rolf ...
Cached Biology News:Mobilizing white blood cells to the lung: New discovery could lead to an improved influenza vaccine 2Can microorganisms be a solution to the world's energy problems? 2Can microorganisms be a solution to the world's energy problems? 3Can microorganisms be a solution to the world's energy problems? 4
... Detection Kit,provides a fast, convenient and ... pyrophosphate (PPi ) produced,by a number ... and,RNA polymerizations, cyclic AMP formation by ... activation of fatty acids to,form their ...
... high PCR specificity *High PCR specificity ... enzyme with unique QIAGEN PCR Buffer ... provided by the stringent built-in hot ... Master Mix and simple reaction setup ...
Request Info...
... AC is the easiest to program ... provided with liquid level sensors for ... integrated orbital shaker and automatic rinsing ... performance. The big bottle model is ...
Biology Products: