Pcr device for qualitative or quantitative detection of nucleic. Lyophilized mix of primers and probes for a total of 96 bcrabl and abl1 reactions each 20 l volume. Product description myco r eal pneumocystis is a realtime pcr assay for detection of dna. For best results, use this master mix with lightcycler 480 multiwell plates. The ideal combination of speed, accuracy, and versatility. The platebased lightcycler 480 system is a highly adaptable and versatile realtime pcr system for gene expression and genetic variation analysis. The cycling conditions for the lightcycler 480 instrument ii are identical. Roche lightcycler 480 instrument setup instructions for qbiomarker copy number pcr arrays. This document is intended as a guide to running kasptm genotyping reactions on the roche lightcycler 480 lc480 instrument. Guide to running kasptm genotyping reactions on the roche. N the performance of the kit as described in this instruction manual is war ranted only when it is used with. Product use limitations the ssoadvanced universal sybr green supermix is intended for research use only, and is not intended for clinical or diagnostic use. Ra lightcycler 480 ii nus yong loo lin school of medicine. See your instrument users manual for detailed instructions on how to load and run the plate.
Lightcycler 480 ii the lightcycler 480 ii instrument is used for performing rapid, accurate polymerase chain reaction pcr in combination with realtime, online detection enabling quantification of a target nucleic acid, as well as postpcr analysis of the amplified nucleic acid by melting curve analysis. Wear appropriate protective eyewear, clothing, and gloves. Setup block type reaction volume l 96 384 20 10 detection format excitation filter emission filter customized format for lightcycler 480 instrument i. Lightcycler 480 realtime pcr system rapid by nature. This test was developed and validated for the lightcycler 1.
However, roche diagnostics gmbh reserves the right to make any changes necessary without notice as part of ongoing product development. However, roche diagnostics gmbh reserves the right to make any changes. The roche lightcycler 480 system with 384well plate thermal block and windows xp data station roche applied science, mannheim, germany was used with barcodelabeled sealed pcr plates to prevent evaporation and contamination. However, it may be used in other types of pcr on the lightcycler 480 system. Primer and probe sequences, as well as optimized concentrations are shown in table below. The lightcycler 480 system utilizes a novel block technology to allow the rapid heating and cooling required to complete a 40cycle, 384well qpcr run in less than 40 minutes. Intended use these instructions are intended for use of eurofins genescan lr, nr, umm kits on roche lightcycler 480 i and ii. The following table shows the pcr parameters that must be programmed for a lightcycler 480 pcr run with the lightcycler 480 sybr green i master. Information specific for single kits can be found in the individual manuals for the respective kits. Taqman fast advanced master mix on the roche lightcycler.
Executed on roche lightcycler 480 realtime pcr system for safety and biohazard guidelines, refer to the safety section in the fast sybr green master mix protocol pn 4385372. For optimal results, select a product length of 500 bp or less. For details on how to program the experimental protocol, see the lightcycler 480 operator. Lightcycler 480 instrument operators manual author. The lightcycler 480 system includes reagents that have been optimized for exceptional sensitivity and specificity in this rapid cycling environment.
The user can create studies that summarize results for multiple experiments i. Pcr instruments, and with the roche lightcycler lc480, qiagen rotorgene q, eppendorf mastercycler ep realplex, and stratagene mx realtime pcr systems. Please follow the procedures in the manual provided with each respective instrument. Identification of the sarscov2 virus occurs by the use of target specific primers and.
Print bookmark share pdf 1602kb english format file size language download get adobe reader contact qiagen. This application manual serves as a general introduction into the principles of realtime pcr and as a guide to the highly diverse applications of this method in life science research. Roboseal adhesive sealing device from hjbioanalytik gmbh d41068 monchengladbach. The readytouse lightcycler 480 sybr green i master offers convenience and ease of use because. Kasp chemistry for allelic discrimination performs well on a roche lc480series machine and this stepbystep protocol will enable users to successfully setup, run and read plates on the lc480. Make sure the correct thermal block cycler 96well for f plate or 384well block format for g plate is in place. Fam red 610 483 558 533 610 customized format for lightcycler 480 instrument ii. But i didnt manage to use it, because the pcr apparatus is a lightcycler 480. The amplicon size should not exceed 750 bp in length.
Creating a color compensation file for roche lightcycler 480 vers. Klasifikace 67548ehs, 199945es neni nebezpecnou latkou nebo smesi. Read the msds and follow the handling instructions. Easytouse hot start reaction mix for pcr using the lightcycler 480 system. The lightcycler 480 ii with led is suitable for detection systems with different probe types such as hybridization probes or hydrolysis 5nuclease taqman probes. See the lightcycler 480 operators manual for general recommendations. N the performance of the kit as described in this instruction manual is war. Along with the primers i added 3 black hole quencher probe and 2x lightcycler probesmaster. With the lightcycler 480 multiple plate analysis software, the user can combine and compare analysis results of lightcycler 480 software 1.
Open the experiment you want to analyze in the main window. Rubella virus real time rtpcr kit user manual for research. Benefit from cuttingedge thermal block and optical technology that leads to excellent welltowell homogeneity and maximized detection. Product characteristics and specifications instruments. The benchtop instrument is easily customizable to meet changing user requirements and can be integrated into everyday use as an automated highthroughput solution.
Lightcycler 480 instrument operators manual is correct at the time of printing. The kit described in this instruction manual has been developed for the lightcycler carouselbased system and the lightcycler 480 system ii 96well block type. All components must be stored at lightcycler 480 ii the lightcycler 480 ii instrument is used for performing rapid, accurate polymerase chain reaction pcr in combination with realtime, online detection enabling quantification of a target nucleic acid, as well as postpcr analysis of the amplified nucleic acid by melting curve analysis. The modern instrument design, outstanding technical and software features, as well as advanced reagents and disposables of the lightcycler 480 system, exceed the demands for fast. The lightcycler 480 system is a proven highperformance, mediumto highthroughput pcr platform that provides various methods for gene detection, gene expression analysis, genetic variation analysis, and array data validation. Roche lightcycler 480 realtime pcr run setup instructions for rt2 profiler pcr arrays. For the applied biosystems 7300, 7500, and 7500 fast realtime pcr.
See your instruments user manual for detailed instructions on how to configure the run method. Automated sample preparation for infectious disease testing. The lightcycler 480 probes master is a readytouse hotstart reaction mix designed specifically for detecting dna targets with hydrolysis probes during lightcycler 480 system pcr. General precautions the user should always pay attention to the following. For optimal results, select an amplicon of 500 bp or less.
Analysis sensitivity depends on the sample volume, elution volume, nucleic acid extraction methods and other factors. The following instructions are based on lightcycler 480 software version 1. Second edition cold spring harbour laboratory part 7 and 8. Benefit from cuttingedge thermal block and optical technology that leads to excellent welltowell homogeneity and. If you use the rna extraction kits recommended, the analysis. Roche lightcycler 480 instrument setup instructions for qbiomarker. Protocol for use with the lightcycler 480 instruments i and ii multiwell plate. Additional equipment and reagents required lightcycler carouselbased system lightcycler 1.
Crossing point cp values cp is equivalent to ct are used to determine unknown sample concentrations from a standard curve. The lightcycler 480 instrument is a versatile, platebased realtime pcr device that supports mono or multicolor. Start the lightcycler 480 software by doubleclicking the icon. All components must be stored at lightcycler 480 qc kit verze 1. Roche lightcycler 480 realtime pcr run setup instructions. Lightcycler 480 instrument operators manual keywords. Setup detection format block type reaction volume sybr green 384 3 20 l programs. The lightcycler 480 realtime pcr system can detect this dye, but it will not be used for fluorescence normalization in the lightcycler 480 system software. Convenient manual analysis tools are implemented in the basic. Taqman fast advanced master mix on the roche lightcycler 480. First presented in 2005, the lightcycler 480 realtime pcr system extends lightcycler system accuracy, speed, and versatility to medium and highthroughput applications in gene expression and genotyping analysis. Lightcycler 480 instrument ii from roche sonoran surplus. The lightcycler carouselbased system provides adapters that allow lightcycler capillaries to be centrifuged in a standard microcentrifuge rotor. Automated sample preparation for infectious disease.
Lightcycler 480, qiagen rotorgene q, biorad cfx96 touch, thermofisher quantstudio 7 pro. Lightcycler 480 96 adjustments may be required for the use of other kits or other realtime pcr instruments. Lightcycler carouselbased instruments and lightcycler 480 instrument. As i read the help pdf file on how to use linreg, i have also dowloaded lc480 converter to convert my text file. Customers using roche in vitro diagnostic lightcycler kits have to use manual a. The lightcycler 480 i system should not be used for this kit. For the applied biosystems 7300, 7500, and 7500 fast realtime pcr systems and the steponeplus realtime pcr systems. Make sure the correct thermal block cycler 96well for f.
Roche applied science lightcycler 480 realtime pcr system. This study is written according to the minimum information for publication of. The use of other reagents has not been tested but is expected to work in the same way. For details on how to program the experimental protocol, see the lightcycler 480 operators manual. Lightcycler 480 ii is a realtime pcr cycler for mid to high sample throughput. Nov 07, 20 i used roches lightcycler 480 to do the experiments. Lightcycler 480 ii lightcycler 480 ii 96 well plate white and plate sealer optical foil 5 transport, storage and stability color compensation kit multiplex 1 is shipped on dry ice or cool packs. Lightcycler 480 realtime pcr system gene quantification. Refer to the operators manual of your realtime pcr instrument for additional.
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