Navigation Links
Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A

Specifically, quantitative real-time RT-PCR on the iCycler iQ was performed in duplicate or triplicate on 1 l of template cDNA per 20 l reaction. Mix A reactions consisted of PCR buffer (16.6 mM (NH4)2SO4, 67 mM Tris, pH 8.8, 6.7 mM MgCl2, 10 mM β-mercaptoethanol; Loging et al. 2000), 1 mM dNTPs (Invitrogen), 0.5 U of Platinum Taq DNA polymerase (Invitrogen), 10 nM fluorescein calibration dye (Bio-Rad), 1 l of a 1:1,500 dilution of 10,000x SYBR Green I stock, 500 nM of each GSP, and 1 l of cDNA. iQ supermix reactions consisted of iQ supermix (Bio-Rad) at a final concentration of 1x, 10 nM fluorescein calibration dye, 1 l of a 1:1,500 dilution of 10,000x SYBR Green I stock, 500 nM of each GSP, and 1 l of cDNA. To control for pipetting losses, 19 l of each 20 l reaction was amplified in a 96- well thin-wall PCR plate (Bio-Rad) using the following PCR parameters: 95C for 2 min followed by 50 cycles of 95C for 15 sec, 60C for 15 sec, and 72C for 15 sec. Melt-curve analysis was performed immediately following amplification by increasing the temperature in 0.4C increments starting at 65C for 85 cycles of 10 sec each. The presence of a single PCR product was verified both by the presence of a single melting temperature peak representing a specific product (vs. a nonspecific primer-dimer peak) using iCycler iQ analysis software and by detection of a single band of the expected size on a 4.5% Super AcrylAgarose gel.

Real-time RT-PCR was performed in duplicate or triplicate reactions. Each GSP pair was used with each reaction mix on each of the two different cDNA templates (derived from macula or periphery). Real-time RT-PCR reactions for detection of the endogenous control gene, A
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. Rapid, High Throughput Mutation Discovery Using Fluorescent SSCP on the ABI PRISM 3100 and 3100Avant Genetic Analyzers
2. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
3. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
4. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
5. Real-Time Multiplex PCR from Genomic DNA Using the iCycler iQ Detection System
6. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
7. Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye
8. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
9. Real-Time Immuno-PCR on the iCycler iQ System, Rev A
10. Real-Time PCR/Melt-Curve Analysis: SNP Detection With FRET, Rev A
11. Real-Time PCR: General Considerations, Rev A
Post Your Comments:
(Date:7/21/2014)... (PRWEB) July 21, 2014 ... Workforce Investment Board (WIB) and Las Positas College ... provide technical education and hands-on training for veterans, ... Technology Program at Las Positas is designed ... needed for engineering technician careers, and establishes a ...
(Date:7/21/2014)... , July 21, 2014  Mike Patterson, CEO ... $1.6 million in Series Seed B funding to ... sensors for the life science, defense, intelligence, and ... Partners led the round and was joined by ... Said Patterson , "This round of funding ...
(Date:7/19/2014)... United States Non-Vascular Stents Market ... report, "United States Non-Vascular Stents Market Outlook to ... States Non-Vascular Stents market. The report provides value, ... and average prices (in US dollars) within market ... Metal Biliary Stents (Metal Non-Covered Biliary Stents, Metal ...
(Date:7/18/2014)... Reflecting the importance of the evolution of Houston’s real estate ... founder and president of Customized Real Estate Services , ... approach to the firm’s branding. , According to Ms. ... of his or her commercial clients have grown exponentially over ... in this market to fully embrace the potential for helping ...
Breaking Biology Technology:Engineering Program Established for Veterans 2Engineering Program Established for Veterans 3Global Graphene Manufacturing Leader, Graphene Frontiers, Receives Series Seed B Funding To Assure Production Of The Next Generation Of Biological And Chemical Sensor Technology 2United States Non-Vascular Stents Market Outlook to 2020 | Researchmoz 2United States Non-Vascular Stents Market Outlook to 2020 | Researchmoz 3United States Non-Vascular Stents Market Outlook to 2020 | Researchmoz 4LEED AP Certified ITRA Global / Customized Real Estate Services Introduces New Approach to Its Branding 2
... Contributed by James V. Anderson and Charles L. Guy, The,University of Florida, Institute ... Introduction , ... , Glucose Regulated Proteins) are ER-luminal proteins ... 1 BiPs are associated with the transport and folding process , ...
... Contributed by Alan T. Bakalinsky Department of Food Science and Technology, ... , Abstract , ... cerevisiae were , compared to those of ... supported previous genetic findings that two of the wine strains are aneuploid , ...
... Marie Nguyen and William Strong, Bio-Rad Laboratories, Inc.,6000 James ... , Introduction , ... is an established technique employed for protein separation. However, ... manual steps and several , hours to ...
Cached Biology Technology:A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF 2A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF 3A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF 4A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF 5Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae 2Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae 3Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae 4Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae 5Electrophoretic Karyotypes of Wine Strains of Saccharomyces cerevisiae 6Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 2Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 3Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 4Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 5Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 6Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 7Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 8Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 9Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 10Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 11Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 12Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A 13
(Date:7/21/2014)... Their scruffy beards weren,t ironic, but there are reasons ... the Ice Age. , According to research from the ... liked living in Greater Cincinnati long before it was ... age. A study led by Brooke Crowley, an assistant ... enjoyed the area so much they likely were year-round ...
(Date:7/21/2014)... July 21, 2014 Dysfunction in dopamine signaling ... genes in the brain,s prefrontal cortex and may ... disorders, such as schizophrenia, according to UC Irvine ... in brain cells that receive this neurotransmitter showed ... affect a variety of behavioral and physiological functions ...
(Date:7/21/2014)... and Aqua satellites detected 154 hotspots in areas across ... land fires had increased again following a decline in ... report was far higher than what had been reported ... , The hotspots were scattered in six regencies and ... areas. Smoke and the related haze it creates could ...
Breaking Biology News(10 mins):Mammoth and mastodon behavior was less roam, more stay at home 2Mammoth and mastodon behavior was less roam, more stay at home 3UCI researchers find epigenetic tie to neuropsychiatric disorders 2
... State University and the University of Chicago has uncovered clues ... N4, injects an unusual substance -- an RNA polymerase protein ... The results, which are published in the current issue of ... of the infection strategies used by viruses that attack bacterial ...
... as an homage to achievement in the field of coral ... link between corals of the Pacific and Atlantic. Dr. Ann ... McNeill of the University of Miami named a new species ... some six million years old after renowned coral reef ...
... was managing several ecosystem research projects on Galveston Island when the 2008 hurricane season began in June 2008. Then ... Click here for more information. , ... , , , , ... , , , , ...
Cached Biology News:Researchers study virus with unusual properties 2Researchers study virus with unusual properties 3Isopora or isn't it? 2Dune and dirty: Hurricane teaches lessons through ecosystem research 2Dune and dirty: Hurricane teaches lessons through ecosystem research 3Dune and dirty: Hurricane teaches lessons through ecosystem research 4Dune and dirty: Hurricane teaches lessons through ecosystem research 5Dune and dirty: Hurricane teaches lessons through ecosystem research 6
... offers the most comprehensive, cost-effective solution ... have incorporated unprecedented quality into both ... includes proven detection chemistries and antibodies ... TechMate protocol. It also includes our ...
Human Cell Line Slides...
Mouse Cell Line Slides...
Request Info...
Biology Products: