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:12/22/2014)... (PRWEB) December 22, 2014 Fertility Associates ... Donor Egg Bank USA , a network of more ... Through this partnership, patients have fast access to a ... vitro fertilization, IVF . Fertility Associates of Memphis is ... this fertility treatment option. , Dr. William Kutteh ...
(Date:12/19/2014)... December 19, 2014 Charm Sciences is ... for the Detection of Aflatoxin M1 in raw commingled ... independent third party validation. The peer reviewed report of ... the Institute for Agricultural and Fisheries Research (ILVO-T&V) has ... , Aflatoxin B1, the most toxic aflatoxin and a ...
(Date:12/19/2014)... Naurex Inc., a biopharmaceutical company leveraging its ... the central nervous system, today announced that Norbert ... present at the 33 rd annual J.P. Morgan ... 3:00 p.m. PST on Tuesday, January 13, 2015, at ... Calif. About Naurex Inc. ...
(Date:12/19/2014)... REDWOOD CITY, Calif. , Dec. 19, 2014 ... , USA) announced today that they have been ... Technologies 1,2 is a privately held company that ... processing of next generation sequencing (NGS) data for the ... into the Roche Sequencing Unit, and will continue to ...
Breaking Biology Technology:Fertility Associates of Memphis Announces Partnership with Donor Egg Bank USA 2Fertility Associates of Memphis Announces Partnership with Donor Egg Bank USA 3Charm Sciences Achieves Independent Validation of First 15 Minute Quantitative Screening Method for Aflatoxin M1 2Naurex to Present at 33rd Annual J.P. Morgan Healthcare Conference 2Bina Technologies is acquired by Roche. 2Bina Technologies is acquired by Roche. 3
... to tight integration and ... excellent value, ... a,leader in radiology and medical image and information management solutions,today announced ... 100 facilities that use MEDITECH(TM) as their health,information system. AMICAS will ...
... May 27 /Xinhua-PRNewswire/ -- Sinovac Biotech Ltd. (Amex:,SVA), ... announced that the,Ministry of Health has purchased approximately ... hepatitis A vaccine, Healive(R), to,vaccinate people living in ... has delivered shipments of the Healive vaccine to,Sichuan, ...
... Calif., May 27 Epitomics, Inc., the rabbit,monoclonal ... its,1000th new rabbit monoclonal antibody. Each antibody has ... proprietary rabbit monoclonal,antibody technology. "We are very excited ... of time. This demonstrated the robustness of our ...
Cached Biology Technology:AMICAS Announces Integrations at More Than 100 MEDITECH Facilities 2AMICAS Announces Integrations at More Than 100 MEDITECH Facilities 3Sinovac Receives RMB 20 Million Purchase Order for Healive from China's Ministry of Health to Vaccinate Earthquake Victims 2Epitomics Announces the Creation of 1000th Rabbit Monoclonal Antibody 2
(Date:12/10/2014)... 2014 You,ve been here before: you desperately need ... your password, site key or the answer to your secret ... teacher? Today, Hoyos Labs , a ... will finally put an end to the frustration that comes ... . 1U leverages a user,s smartphone to acquire his or ...
(Date:12/10/2014)... , December 9, 2014 ... BCARD Platform   ... meeting scheduling solutions for business-to-business (B2B) events, today ... leader in mobile near-field communication (NFC), Bluetooth low ...      (Logo: http://photos.prnewswire.com/prnh/20140612/691055) , Jifflenow will ...
(Date:12/3/2014)... 2014 As part of our commitment to ... pleased to announce the release of a new reader ... the workforce data that they need. The ... by existing readers. Many such devices have serious shortcomings ... technology. Older models force users to navigate numerous complicated ...
Breaking Biology News(10 mins):The Password is Finally Dead: Launch of 1U Mobile App Eliminates Need for All Usernames and Passwords 2The Password is Finally Dead: Launch of 1U Mobile App Eliminates Need for All Usernames and Passwords 3Jifflenow And ITN International Bring Cutting-Edge Badge Scanning Technology To B2B Events 2Jifflenow And ITN International Bring Cutting-Edge Badge Scanning Technology To B2B Events 3Inception Technologies to Release New Biometric Reader 2
... enzyme Akt1 could provide the key to boosting the ... dendritic cells, the immune-system master switches that promote the ... Baylor College of Medicine in a report that appears ... issue of the journal Nature Biotechnology. , "By keeping ...
... de Investigaciones Científicas (CSIC) has discovered a new route ... discovery that will enable the biotechnological design of plant ... produce oral vaccines which, upon being ingested, will be ... the door to the design of protein-manufacturing plants of ...
... Bioinformatics Institute and Duke University Medical Center have ... an important role in the development and virulence ... fungal pathogen that causes black spot disease on ... leaf loss in many economically important crops including ...
Cached Biology News:'Super' enzyme may lead way to better tumor vaccines 2Vaccine-producing 'plant-factories' 2Tearing down the fungal cell wall 2Tearing down the fungal cell wall 3
... Disrupts biological cells for DNA ... 96 well plates (includes 96 well ... consistent lysis across the entire plate ... wells). Can be used for ...
Rat polyclonal to acetyl Salicylic Acid ( Abpromise for all tested applications)....
Rabbit polyclonal to UBC12 (rating: ****) ( Abpromise for all tested applications). Antigen: Synthetic peptide: RGGYIGSTYFER, corresponding to amino acids 169-180 of UBC12 Entrez Gene ID: 90...
...
Biology Products: