What is LightCycler 480?
The LightCycler® 480 System is a proven high-performance, medium-to high-throughput PCR platform that provides various methods for gene detection, gene expression analysis, genetic variation analysis, and array data validation.
What is LightCycler?
LightCycler technology combines rapid-cycle polymerase chain reaction with real-time fluorescent monitoring and melting curve analysis. Since its introduction in 1997, it is now used in many areas of molecular pathology, including oncology (solid tumors and hematopathology), inherited disease, and infectious disease.
What is Roche LightCycler?
The LightCycler® 96 Instrument is a fast and precise 96-well real-time PCR device. It provides various methods for gene expression and genetic variation analysis in mono- or multicolor (up to 4 colors*) mode. The software includes analysis modules for: Relative Quantification. Absolute Quantification.
Why is quantitative PCR called real time PCR?
The fluorometer detects that fluorescence in real time as the thermal cycler runs, giving readings throughout the amplification process of the PCR. As a result, quantitative PCR is also called real-time PCR or RT-PCR.
Why is a thermal cycler used in PCR?
Thermocyclers, or thermal cyclers, are instruments used to amplify DNA and RNA samples by the polymerase chain reaction. The thermocycler raises and lowers the temperature of the samples in a holding block in discrete, pre-programmed steps, allowing for denaturation and reannealing of samples with various reagents.
Why is quantitative PCR called real-time PCR?
What are the applications of qPCR?
Multiple research applications based on qPCR have been developed. These include gene expression analysis, genotyping, allelic discrimination, single nucleotide polymorphism (SNP) detection, microbe and pathogen detection, species abundance quantification and many others.
What is qPCR used for?
qPCR stands for quantitative polymerase chain reaction and is a technology used for measuring DNA using PCR.
Where are thermal cyclers used?
Thermal cyclers are used in a range of disciplines. The majority of thermocyclers reside in research laboratories, where they are mainly used for amplifying DNA. In addition, researchers take advantage of thermal cyclers for cloning DNA and sequencing genes.
What can thermocycler be used for?
Thermocycler Use in Life Science Research
In molecular biology they are used for DNA sequencing, cloning, generation of probes, quantification of DNA and RNA, studying patterns of gene expression, detection of sequence-tagged sites, and many more techniques.
What is real-time PCR and its application?
Real-time PCR is a widely used technology for mutation detection that not only provides excellent sensitivity and specificity when paired with our gold standard TaqMan Assays, but also provides rapid results and is easy to perform when compared to other methods.
What is real-time PCR used for?
Real-time PCR/qPCR assays have become the tool of choice for the rapid and sensitive determination and quantitation of nucleic acid in various biological samples, with diverse applications such as gene expression analysis, the detection of genetically modified organisms in food, and cancer phenotyping.
What does a melt curve allow you to do?
Researchers often use melt curve analysis to assess whether their intercalating dye PCR/qPCR assays have produced single, specific products.
What are the most commonly used methods used to detect products in qPCR?
Quantitative PCR
The two common methods used to detect and quantify the product include (1) fluorescent dyes that non-specifically intercalate with double-stranded DNA and (2) sequence-specific DNA probes consisting of fluorescently labeled reports.
What is the application of the thermal cycler in PCR?
The Thermal Cycler (also known as a Thermocycler, PCR Machine or DNA Amplifier) is a laboratory apparatus used to amplify segments of DNA via the Polymerase Chain Reaction (PCR). The device has a thermal block with holes where tubes holding the PCR reaction mixtures can be inserted.
What is a PCR machine used for?
PCR-machines are used for replication of DNA, detecting DNA sequences, carrying out DNA fingerprinting, forensic analysis, and molecular cloning, diagnosing genetic diseases, and detecting pathogens such as Hepatitis B and C viruses, Chlamydia trachomatis, HIV-1 causing AIDS, Neisseria gonorrhoeae, human papillomavirus …
What is RT-qPCR used for?
The final acronym ‘RT-qPCR’ is used for reverse transcription quantitative real-time PCR. This is a technique which combines RT-PCR with qPCR to enable the measurement of RNA levels through the use of cDNA in a qPCR reaction, thus allowing rapid detection of gene expression changes (see Figure 1C).
What does a qPCR melt curve show?
A typical denaturation (melt) curve performed after qPCR cycling with an intercalating dye, will typically give rise to a single distinct peak in the plot of the negative derivative of fluorescence vs. temperature. This indicates that the amplified double-stranded DNA products are a single discrete species.
How does real time qPCR work?
In the one-step method, RT and qPCR are performed in the same tube. A real-time PCR detection system consists of a thermal cycler equipped with an optical detection module to measure the fluorescence signal generated during each amplification cycle as the fluorophore binds to the target sequence.
What are the applications of polymerase chain reaction?
We present a survey of the following applications of PCR: 1) The amplification of gene fragments as fast alternative of cloning. 2) The modification of DNA fragments. 3) The sensitive detection of pathogenic microorganisms, if desired followed by an accurate genotyping. 4) DNA analysis of arachaeological specimens.
What are applications of real-time PCR?
What are thermal cyclers used for?
What is the difference between RT-PCR and qPCR?
1. QPCR and RT-PCR are both terms used in biotechnology and utilized for the production of multiple copies of DNA. 2. RT-PCR is used to amplify the reversed transcription of the DNA code; QPCR measures the amplification.
What is melt curve analysis used for?
Melt curve analysis is frequently used as a diagnostic tool for assessing qPCR amplicon length with intercalating dye qPCR assays. Here, we explain how melt curves are produced, examine the assumptions being used, and describe some additional methods that can be used to further analyze melt curve results.
Can you do a melt curve with Taqman probes?
Taqman probe can’t be used for melting curve analysis. Only when using Sybr-Green or similar intercalating dyes need to do melting curve analysis as Sybr-green is non-specific bind to double stranded DNA. Taqman is specific bind to template sequence.