New from Roche: The LightCycler<sup>®</sup> 480 High Resolution Melting Master for gene scanning
3 Apr 2007High Resolution Melting (HRM) is a novel, close-tube, post-PCR method enabling genomic researchers to analyze genetic variations (SNPs, mutations, methylations) in PCR amplicons prior to or as an alternative to sequencing. It provides high specificity, sensitivity and convenience at significantly higher speed and lower cost much than other established (e.g., gel- and/or gradient based) methods (e.g., SSCP, dHPLC).
The LightCycler® 480 High Resolution Melting Master contains a special generic DNA dye which binds to DNA in a saturating fashion and without preference for certain regions or conformations. It gives rise to very sharp and distinct melting signals which, when acquired at high resolution, can reveal very subtle differences between e.g., wild-type and heterozygous samples. The dye contained in the LightCycler® 480 High Resolution Melting Master is novel and proprietary and chemically different from other HRM dyes like LC Green Plus (Idaho Tech) or SYTO9 (Invitrogen). It works at much lower concentrations (~ten fold) than other dyes and can therefore be used in a PCR master mix without interfering with PCR performance. It also shows improved stability and signal dynamics and gives rise to melting curves whose shape can be easily analyzed and interpreted
The main application for the HRM method is gene scanning – the search for unknown genetic variations (SNPs, mutations, deletions) in PCR amplicons (the detection and calling of already known mutations is referred to as genotypin). Gene scanning on the LightCycler® 480 System is based on the analysis of melting curve data obtained at high resolution (HRM). With HRM, unknown sequence variations in diploids become apparent in heterozygous samples because they have significantly differently shaped melting curves compared to those derived from homozygous (wildtype or mutant) samples. The LightCycler® 480 Instrument is the first plate-based real-time PCR instrument facilitating such analysis due to its high-performance thermal control, and optical components.
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LightCycler® 480 Real-Time PCR System
The Roche LightCycler 480 System - Another breakthrough in real-time PCR analysis of gene expression and melting curve-based mutation analysis.The Roche LightCycler 480 RT-PCR System combines amazing speed with exceptional accuracy and will meet the needs of a broad range of scientific applications in genomics research including array validation, gene-knockdown studies, and SNP analysis.This Roche Light Cycler PCR supports all current probe formats and applications and its built-in versatility allows adaptation to future technological advances in genomics research.Customer benefits from the Roche LightCycler® 480 System: Speed – rapid temperature control for superb specificity and yield.The LightCycler 480 PCR System utilizes a novel block technology to allow the rapid heating and cooling required to complete a 40-cycle, 384-well quantitative PCR run in less than 40 minutes and leads to extraordinary well-to-well temperature homogeneity and maximized inter-well, inter-cycle reproducibility. Accuracy – unbiased conditions for unambiguous results Special arrangement of optical components ensures the uniform collection of signals across the plate and makes analysis independent of sample position or differences in reaction volume. Versatility – utmost flexibility for maximum efficiency Five excitation and six emission filters of the Roche LightCycler allow you to work with the assay format of your choice, including SYBR Green I, hydrolysis probes, probes, probes, Molecular Beacons, and others. High-performance optical system from the LightCycler 480Fluorescence excitation by a high-intensity xenon lamp, and a specially developed signal-collection system, result in maximal sensitivity for any assay format in all channels. Additional Features of the LightCycler 480 PCR System: Customizable modular system concept Building on the principles of the well-established, performance-proven LightCycler® Software 4.0, the Roche LightCycler® 480 platform comes equipped with modules for run control, Tm calling, and absolute quantification. Further customize your system to your specific needs by adding separately offered advanced analysis modules. Especially developed reagents A tailor-made hot-start PCR enzyme master mix is available for each type of LightCycler® 480 application. Built on Roche Diagnostics’ performance proven FastStart technology, these reagents ensure optimal reproducibility and high dynamic ranges. Perfect integration into laboratory workflow Compatibility for automated sample-preparation, automated loading and connectivity to laboratory information management systems (LIMS) for integrated workflow monitoring offer uncompromising adaptability for a perfect fit.