On this page the most prominent real-time PCR cycler are described. In the cycler descriptions the specifications and the advantages of the displayed systems are shown. Which real-time platform meets your requirements best, depends on your research application. Some of the systems are designed for research with low capacities and others are for high-throughput applications, most in combination with pipetting robots. Most of them use solid-block for thermal cycling, other use hot- and cooled-air. Most differences are obviously in the application software, especially in the way of data analysis and how the derived crossing points or threshold levels are computed. Each of these systems employs either one of several general types of fluorescent probes for detection. There are also big differences how data are displayed. Some of the limitations of end-point detection in (RT-) PCR have been assuaged in real-time PCR systems, a number of which are now on the market. These systems offer many general technical advantages, including reduced probabilities of variability and contamination, as well as online monitoring and the lack of need for post reaction analyses. Further, some of these systems were developed with contemporary applications such as quantitative PCR, multiplexing, and high-throughput analysis in mind. Initial template levels can be calculated by analysing the shape of the curve or by determining when the signal rises above some threshold value. Several commercially available real-time PCR systems are overviewed on this page and/or summarized in the accompanying table. summary in PDF
NEW ! StepOne
Real-Time PCR System
The 9800 Fast PCR System delivers the
high-quality results that you've come to expect from Applied Biosystems
The StepOne™ Real-Time PCR System combines plug and play convenience and uncompromising performance that is tailor-made for both first-time users and real-time PCR experts. By utilizing the full range of TaqMan™ gene expression and genotyping assays, the StepOne System delivers the kind of high-quality results you've come to expect from the leader in real-time PCR. The StepOne System provides the power and flexibility experienced users demand, while its intuitive software guides new users through experimental design, operation, and analysis. Part of an elegant, optimized system solution for real-time PCR that includes software, reagents, and instrumentation, the StepOne System delivers remarkable performance with remarkable ease-of-use, at a price that can only be described as...remarkable. * Easy to use software helps you set up your experiments and get results quickly * Fast runs (<40 minutes) and standard runs (<2 hours) with one block * Flexible configurations for PC-free or networked environments * Preloaded and precalibrated to get you up and running fast * Small, space-saving footprint * Supports real-time PCR applications  Applied Biosystems’
latest introduction to real-time PCR, the StepOnePlus™ Real-Time PCR
System, makes it simple and easy to get high-quality real-time PCR
results. This remarkably simple 96 well, 4 color, real-time PCR system
is designed with a user-friendly, yet powerful, interface for
researchers of all experience levels.
* Intuitive, flexible software and wizards guide new users through their real-time PCR experiments in three easy-to follow steps * 4-color/96-well format delivers precise, quantitative real-time PCR results * Long-life LED-based optical system records fluorescence from FAM™/SYBR® Green, VIC®/JOE™, ROX™ and NED/TAMRA™ dyes for gene expression analysis, pathogen detection, SNP genotyping, and presence/absence assays * One block performs both standard runs in under 2 hours and fast PCR reactions in less than 40 minutes * Ultra-compact footprint fits any laboratory setting * LCD touchscreen and USB drive provide configuration flexibility and enable PC-free operation * Remote monitoring and email notification for convenience and time-savings * VeriFlex™ Block technology brings six independently controlled peltier blocks together for precise temperature control and enhanced PCR functionality * For those customers who currently have a StepOne™ System, an upgrade option will be available soon. Remarkably Simple System The StepOnePlus™ system brings advanced real-time PCR technology to a new level of accessibility. Beginning at the StepOnePlus software homepage, you can seamlessly navigate through all aspects of the real-time PCR method including sample and reaction set-up, thermal cycling, and fluorescent detection. Focused application software analyzes and interprets experimental results. Depending on the experimental design, the system can even help you select and order real-time PCR reagents online by means of convenient links in the Design Wizard. Simply Remarkable Results Because the StepOnePlus system is factory-calibrated for optical and thermal accuracy, simply remarkable real-time PCR results are available right out of the box. It can discriminate between 2 populations of 5,000 and 10,000 template copies of a TaqMan® Assay with 99.7% confidence. Applications and Software Analysis The StepOnePlus System supports any real-time PCR application. * SNP Genotyping * Gene Expression Profiling * MicroRNA Expression * Translocation Analysis * Gene Detection * Viral Load Analysis Software for StepOne and StepOnePlus systems supports a variety of analysis methods, including: * Absolute Quantitation * Standard Curve * Relative Quantitation * Relative Standard Curve * Comparative CT (∆∆ CT) * Presence/absence (Plus/Minus) assays with an internal positive control * Melt curve analysis * Genotyping (including real-time amplification) Software Instrument software for the Applied Biosystems StepOne Real-Time PCR System runs on the Windows XP® operating system and provides instrument control, data collection, and data analysis. This user-friendly and intuitive software package includes the following features: * Experimental design wizards to help you design and set up experiments. * Pipetting protocols and recipes to quickly set up experiments. * Advanced setup for expert users who require flexibility for more complex applications, such as multiplexing. * QuickStart setup so that you start a run immediately and enter plate information at a later time. * Real-time monitoring of amplification growth curves enables you to view run progress. * Remote real-time monitoring so that you can monitor progress from a remote PC. * Email notifications to alert you when a run has started or ended. * Auto-baseline and auto-threshold for simplified data analysis. * Automated SNP genotype calling capability with intuitive graphical output and quality-value assignment. * Tool tips for easy identification of sample wells when viewing amplification curves or SNP genotyping plots. * Troubleshooting flags to help you diagnose and solve problematic experiments. * Multiple plots view so that you can simultaneously assess data from four different perspectives. * Easy cut and paste functionality. * Export easily to PowerPoint, Excel, and graphical images read more download StepOnePlus brochure Introducing the NEW Applied Biosystems 7500 Fast real-time PCR System
The LightCycler™ from Roche Molecular Biochemicals of
Indianapolis harnesses light technology licensed from Idaho Technology
Inc. of Idaho Falls, and then adds some additional bells and whistles,
such as a built-in microvolume fluorimetric detection system that
enables real-time quantitative PCR. This system employs thin-walled
glass capillary tubes, and typical PCR experiments can be performed in
less than 30 minutes. Currently, this system supports two
fluorescence-based methods for the detection of amplification products:
the "general" DNA stain SYBR Green I (a product of Molecular Probes of
Eugene, Ore.), or sequence-specific hybridization probe pairs. SYBR
Green I exhibits very little fluorescence when free in solution;
emission is greatly enhanced when it binds to the minor groove of the
DNA double helix. Prior to amplification, the reaction mixture contains
the denatured DNA, the primers, and the dye. The low-level background
fluorescence signal generated by the unbound dye molecules is
subtracted during computer analysis. After annealing of the primers, a
few dye molecules can bind to the double strand. During elongation,
more and more dye molecules bind to the newly synthesized DNA,
resulting in dramatically increased light emission. If the reaction is
monitored continuously, this increase in fluorescence can be viewed in
real time. After denaturation of the DNA during the next heating cycle,
the dye molecules are released and the fluorescence signal falls. A
fluorescence measurement is performed at the end of the elongation step
of every PCR cycle to monitor the increasing amount of amplified DNA.
The hybridization probe format employs two specially designed,
sequence-specificoligonucleotides lab-eled with fluorescent dyes. One
oligonucleotide probe carries a fluorescein label at its 3' end;
the other probe carries a different label (LC Red 640 or LC Red 705) at
its 5' end. The chemical nature of the hybridization probes prevents
their extension: one probe contains fluorescein at the 3' end, whereas
the 5'-labeled probe contains a 3' phosphate moiety. The sequences of
the two oligonucleotides are selected so that they hybridize to the
amplified DNA fragment in a head-to-tail arrangement. When the
oligonucleotides hybridize in this orientation, the two fluorescent
dyes are positioned in close proximity to each other. The first dye
(fluorescein) is excited by the LightCycler's light emitting diode
(LED) filtered light source, and emits green fluorescent light at a
slightly longer wavelength. When the two dyes are in close proximity,
the emitted energy excites the dye attached to the second hybridization
probe, which subsequently emits red fluorescent light at an even longer
wavelength. This energy transfer, referred to as fluorescence resonance
energy transfer (FRET), occurs efficiently only when the dyes are in
close proximity (a distance between 1-5 nucleotides). Thus, in this
type of assay, fluorescent intensity measurements are made after the
annealing steps. The increasing amount of emitted fluorescence is
proportional to the increasing amount of DNA generated during the
linear phase of the ongoing PCR process.
by Idaho Technology http://www.idahotec.com/rapidcycler/index.html
The RapidCycler is the first instrument engineered to match the speed of biochemical reactions. The instrument's rapid temperature cycling system is based on heat transfer by hot air to samples contained in microcapillary tubes or thin walled micorcentrifuge tubes. Heating and cooling the samples with blasts of high velocity air results in nearly instantaneous temperature transactions, ensures temperature uniformity and rapid heat exchange within the sample.
|
||||||||||||||||
Mastercycler® ep realplex - real-time PCR solutions
|
