Navigation Links
Identification of Nonspecific Products Using Melt-Curve Analysis on the iCycler iQ Detection System, Rev A

Marni Brisson, Larissa Tan, Rob Park, and Keith Hamby, Bio-Rad Laboratories, Inc., 2000 Alfred Nobel Dr., Hercules, CA 94547 USA


Introduction
Real-time PCR* is a powerful and effective technique for accurate quantitation of DNA. Assays for the detection of a single gene involve careful choice of primers, target sequence, and the method for detecting the amplified DNA product. In addition, it is necessary to choose appropriate reaction conditions that will generate robust and efficient amplification for both relative and absolute quantitation.

Specificity and efficiency are two essential requirements for successful real-time PCR amplification. Poor assay design can result in such problems as nonspecific products and inefficient primer association with target, which dramatically affect the quality of the data. Proper primer design ensures that the chosen primers are specific for the desired target sequence, bind at positions that avoid secondary structure, and minimize the occurrence of primer-dimer formation. Appropriate reaction conditions can also improve the efficiency of amplification. Optimization of the detection method, Mg2+ concentration, annealing temperature, enzyme concentration, PCR product length, etc., are all ways of perfecting real-time assays.

Another valuable tool in assay design is melt-curve analysis. With the use of DNA-binding dyes such as SYBR Green I, a melt-curve profile can be generated. The iCycler iQ system records the total fluorescence generated by SYBR Green I binding to double-stranded DNA as temperature changes, and plots the fluorescence in real time as a function of temperature. The first derivative of this plot, dF/dT, is the
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12

Related biology technology :

1. Identification of Differentially Expressed Gene Products with the CastAway System*
2. An Optimal DNA Microarray Substrate for the Identification of Fetal and Somatic Liver Stem Cells of the Rat
3. Identification of Low-Level Oxidation Products in Calmodulin Using NanoSpray Ion Trap Mass Spectrometry
4. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
5. Protect RNA and Improve Target Cell Identification
6. Identification of Disease Causing Mutations in Phenylketonuria by Denaturing Gradient Gel Electrophoresis Using the DCode System
7. Identification of cells in S phase using the Cell Proliferation Fluorescence Kit and IN Cell Analyzer 1000
8. Identification of Insecticides and Herbicides by LC/MS/MS Using the API 2000 LC/MS/MS System
9. Identification of Phase I and Phase II Metabolites of Buspirone on the Q TRAP LC/MS/MS System
10. ESR1 Gene Expression in Oncology and Metabolic Diseases Using the ASCENTATM System Target Validation and Identification of Novel Disease Indications
11. High-Throughput System Generates Ultra-Pure PCR Products Suitable for All Applications
Post Your Comments:
(Date:7/28/2014)... Research and Markets has announced the ... report to their offering. ... technique used to determine the size of a particle in ... to analyze the size of particles as it can affect ... performed to determine the behavior and nature of a material, ...
(Date:7/25/2014)... 2014 "Ischemic Stroke and Cerebral ... data on the Ischemic Stroke and Cerebral Stroke ... and data relating to the clinical trials on ... global Ischemic Stroke Research Report 2014 at: ... the trial numbers and their recruitment status as ...
(Date:7/25/2014)... by researchers from the University of Leicester has ... unlocking the potential to create new materials using ... been published in the prestigious academic journal ... technique to examine in rich detail the structure ... containing an acetylene molecule and a single helium ...
(Date:7/25/2014)... 2014 WABC Radio show out of New ... Lorry Young will be hosting California-based Vet-Stem, Inc.’s Founder and ... therapy in pets. Dr. Harman first visited the show ... therapy for pets suffering from osteoarthritis and other degenerative diseases, ... industry forward. , Young has invited Dr. Harman back to ...
Breaking Biology Technology:Global Particle Size Analysis Market 2014-2018 2Ischemic Stroke and Cerebral Stroke Global Clinical Trials Review H2, 2014: MarketResearchReports.Biz 2Ischemic Stroke and Cerebral Stroke Global Clinical Trials Review H2, 2014: MarketResearchReports.Biz 3Breakthrough laser experiment reveals liquid-like motion of atoms in an ultra-cold cluster 2Breakthrough laser experiment reveals liquid-like motion of atoms in an ultra-cold cluster 3Robert J. Harman, DVM, Founder and CEO of Vet-Stem, Inc. to Join New York Radio Host Lorry Young in Upcoming Episode of A “Paws” For Your Pet 2Robert J. Harman, DVM, Founder and CEO of Vet-Stem, Inc. to Join New York Radio Host Lorry Young in Upcoming Episode of A “Paws” For Your Pet 3
... Productivity. , ... Grand Forks, ND (Vocus) December 10, ... individual management practices and fields will become increasingly important to growers, ... Research and Development, will tell agricultural leaders today at the ...
... Dosing Regimen , , ... long-acting therapy (LAT) demonstrated statistically significant symptom control compared ... study presented today (1). Statistical significance was evidenced ... equivalent [eq.a]), when given every 4 weeks with a ...
... Journal Blood for the American Society of Hematology Annual Meeting ... 10 Alfacell Corporation (Nasdaq: ACEL ) today ... in Blood (2008 112: Abstract 4205 & 2008 ... show anti-tumor activity in chronic lymphocytic leukemia (CLL) and acute ...
Cached Biology Technology:Matching Genetics, Technology and Management Practices Critical to Meeting Strong Demand for Agricultural Crops 2Matching Genetics, Technology and Management Practices Critical to Meeting Strong Demand for Agricultural Crops 3Matching Genetics, Technology and Management Practices Critical to Meeting Strong Demand for Agricultural Crops 4Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 2Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 3Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 4Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 5Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 6Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 7Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 8Phase 3 Data Demonstrate Efficacy and Tolerability of Paliperidone Palmitate, an Investigational Long-Acting Therapy for the Treatment of Schizophrenia 9Alfacell's ONCONASE(R) and R-Amphinase Show Anti-Tumor Activity in Chronic Lymphocytic Leukemia and Acute Myeloblastic Leukemia Cells 2Alfacell's ONCONASE(R) and R-Amphinase Show Anti-Tumor Activity in Chronic Lymphocytic Leukemia and Acute Myeloblastic Leukemia Cells 3Alfacell's ONCONASE(R) and R-Amphinase Show Anti-Tumor Activity in Chronic Lymphocytic Leukemia and Acute Myeloblastic Leukemia Cells 4Alfacell's ONCONASE(R) and R-Amphinase Show Anti-Tumor Activity in Chronic Lymphocytic Leukemia and Acute Myeloblastic Leukemia Cells 5
(Date:7/28/2014)... Eight of the top 10 U.S. cities that have ... public inconveniences as frequent road closures, overwhelmed storm drains ... a new NOAA technical report. , This nuisance flooding, ... three U.S. coasts, between 300 and 925 percent since ... Nuisance Flood Frequency Changes around the United States, also ...
(Date:7/28/2014)... study suggest two ways to effectively address the problem that ... women, compared to women of a normal body mass index. ... but as the population has increased in weight, concern has ... Studies have consistently found that obesity has a negative impact ... affect how well the pill prevents pregnancy. , ...
(Date:7/28/2014)... He calls himself the bug hunter, but the ... only be found and identified with special methods and ... Kansas State Veterinary Diagnostic Laboratory at Kansas State University, ... porcine enterovirus G, which is an important find in ... in cells, but didn,t know what it was," Hause ...
Breaking Biology News(10 mins):NOAA: 'Nuisance flooding' an increasing problem as coastal sea levels rise 2NOAA: 'Nuisance flooding' an increasing problem as coastal sea levels rise 3Strategies identified to improve oral contraceptive success with obese women 2Researcher using next-generation sequencing to rapidly identify pathogens 2Researcher using next-generation sequencing to rapidly identify pathogens 3
... Junrong Zheng,s techniques to see the fine details of how ... highly prestigious Packard Fellowship. Zheng and his team ... grant that comes with the fellowship to build a laser-based ... and orientation -- of any molecule, no matter how complex. ...
... BOSTON, MA (October 20, 2011) Results from a ... copper surfaces in intensive care unit rooms reduced the amount ... in a 41 percent reduction in the hospital acquired infection ... first to demonstrate the value of a passive infection control ...
... of Bristol has been awarded 1.3 million to unlock the ... to produce silica the fundamental constituent of glass. It ... the next generation of medical imaging tools. Inspired by ... Paul Curnow from the University,s School of Biochemistry has been ...
Cached Biology News:Rice's Zheng wins Packard Fellowship 2Study proves new technology kills bacteria 2
PTC-100 cycler with gold/silver block with Hot Bonnet heated lid, holds one 96-well V-plate or96x0.2ml tubes...
... RNA (bluish-purple), DNA (blue), proteins (red) and acid polysaccharides ... Cation traces: Ca ≤ 10 mg/kg Cd ≤ 5 ... Cu ≤ 5 mg/kg Fe ≤ 5 mg/kg K ... 5 mg/kg Na ≤ 50 mg/kg Ni ≤ 5 ...
... The DTX Series Multimode Detectors provide ... variety of application and automation needs. ... absorbance measurements on the same instrument, ... standalone instruments or can be integrated ...
BioGenex IHC Kits for Rodent Tissues are designed for immunohistochemistry on rodent and rabbit tissues that are closely related to the source of the primary antibody. This kit comes with rat seconda...
Biology Products: