Readytouse hot start reaction mix for sybr green ibased realtime pcr using the lightcycler 480 instrument. Lightcycler 480 software setup for the quantifast sybr. Powerup sybr green master mix thermo fisher scientific us. Brilliant iii ultrafast sybr green qpcr master mix quick. When used with the lightcycler 480 system, when used with the lightcycler 480 system, this kit is ideally suited for hot start pcr applications. Lightcycler lc480, qiagen rotorgene q, eppendorf mastercycler ep realplex, and stratagene mx realtime pcr systems. Hybridization probe that detects gene of interest consisting of a primer pair with a 3. Lightcycler 480 software setup for the quantifast sybr green rtpcr kit en. Introducing the lightcycler 480 system the next generation of. Lightcycler 480 sybr green i master contains components which help to minimize. The cycling conditions we used were optimized for the roche kit. Wear appropriate protective eyewear, clothing, and gloves. Sybr green master mixes on the roche lightcycler 480 real. Reaction mix preparation and thermal cycling protocol 1.
As i read the help pdf file on how to use linreg, i. The lightcycler 480 sybr green i master is a onecomponent hot start reaction mix for pcr. Thaw ssoadvanced universal sybr green supermix and other frozen reaction components to room temperature. The kit is designed for 96 or 480 reactions depending on pack size, with a. It contains antibodymediated hotstart itaq dna polymerase, dntps, mgcl 2, sybr green i dye, enhancers, stabilizers, and a blend of. Lightcycler 480 sybr green i master mix roche diagnostics. Make dilutions of cdna or dna to be used for the standard curve. Typical sybr green reaction protocol reaction setup reagent final concentration volume l one reaction water 1. Lightcycler faststart dna master sybr green i readytouse hot start reaction mix for pcr using the lightcycler1 instrument cat. 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. Kits contain primers, dna standards, and qpcr reagents specific for both dna sequencing platform and qpcr instrument.
During each phase of dna synthesis, the sybr green i dye, which is included in the reaction mix, binds to the amplified pcr products. Lightcycler 480 sybr green i master and probes master available in pack sizes suitable for highthroughput applications. Enter sample information for your experiment as needed. Lightcycler 480 sybr green i master mix roche diagnostics 2. Powerup sybr green master mix thermo fisher scientific kr. Lightcycler 480 sybr green i master using the lightcycler 480 system include. For in vitro lightcycler faststart dna master sybr green i. Absolute quantification techniques are used to determine the amount of target dna in the initial sample, while relative quantification determines the ratio between the amount of target dna and a reference amplicon. In combination with the lightcycler 480 system and suit able pcr primers, this kit allows very sensitive detection and quantification of. Roche diagnostics lightcycler 480 sybr green i m lightcycler. Taqman fast advanced master mix on the roche lightcycler. To save the report as a pdf file, click the pdf button in the tool bar. Sybr green qpcr methods applied written by posted on.
Maxima sybr green qpcr master mixes are compatible with most realtime thermal cyclers. Perfecta sybr green fastmix is a 2x concentrated, readytouse reaction cocktail that contains all components, except primers and template for realtime quantitative pcr systems that do not require. Brilliant iii ultrafast sybr green qrtpcr master mix. Taqman wikipedia bioinformatics pcr efficiency in realtime pcr full. Lightcycler 480 system reagents roche life science. Read the msds and follow the handling instructions. Description gotaq qpcr master mixa,b is a reagent system for quantitative pcr qpcr. The complete pcr mix that is lightcycler 480 sybr green master supplemented with primers and template is. Exposure to direct light for an extended period of. Is anyone using life technologies sybr green master mix on a. Lightcycler 480 sybr green i master is designed for research studies. Lightcycler rna amplification kit sybr green i cat.
Introducing the lightcycler 480 system the next generation. Fast sybr green master mix performance on the lightcycler 480 realtime pcr system. Executed on roche lightcycler 480 realtime pcr system for safety and biohazard guidelines, refer to the safety section in the power sybr green pcr master mix and rtpcr protocol pn 4367218. Brilliant iii ultrafast sybr green qrtpcr master mix quick. Faststart dna master sybr green i is designed for life sience. Easytouse hot start reaction mix for pcr using the lightcycler 480 system. 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. Rapid highthroughput, platebased realtime pcr amplification and detection instrument. The lightcycler faststart dna master sybr green i can also be used to perform a twostep rtpcr protocol. When used with the lightcycler 480 system, this kit is ideally suited for hotstart pcr applications. The kapa sybr fast qpcr kit is supplied as a 2x master mix with. Powerup sybr green master mix can be used in either standard or fast cycling mode and is compatible with all applied biosystems realtime pcr instruments. Power sybr green pcr master mix thermo fisher scientific. Make dilutions of cdna or dna to be used for the standard.
For onestep rtpcr protocol using the lightcycler instrument refer to. It contains faststart taq dna polymerase and dna double strand. Is anyone using life technologies sybr green master mix on. Kapa sybr fast qpcr master mix 2x optimized for the lightcycler 480 is a readytouse cocktail containing all components except primers and template, for the amplification and detection of dna on the lightcycler 480 realtime pcr detection system. Lightcycler 480 instrument operators manual is correct at the time of printing. Contains faststart taq dna polymerase, reaction buffer, dntp mix with dutp instead of dttp, sybr green i dye, and mgcl 2. Turn on the computer linked to the instrument and log on to window xp. The fast sybr green master mix protocol pn 4385372 provides detailed realtime pcr and. Where noted, master mixes contain instrumentspecific reference dyes, while the universal kit includes rox high and rox low both 50x separately. We tested the sensifast sybr norox kit against the lightcycler 480 sybr green i master on a roche lightcycler 480.
The kit is designed for 96 or 480 reactions depending on pack size, with a final reaction volume of 20 l each. One of the following reagents is required for reverse transcription of rna into cdna. The lightcycler 480 system is a highperformance, medium to highthroughput pcr platform 96 or 384well plates that provides various methods for gene detection, gene expression analysis, genetic variation analysis, and array data validation. Enter username and password when the login dialog box appears and click the button to proceed with the login. Lightcycler 480 software setup for the quantifast sybr green. Lightcycler 480 sybr green i master version june 2005 easytouse hotstart reaction mix for pcr using the lightcycler 480 system cat. Lightcycler 480 sybr green i master from roche description readytouse hot start reaction mix for sybr green ibased realtime pcr using the lightcycler 480 instrument. For optimal results, recommended primer concentrations should. The system contains a fluorescent dnabinding dye, the bryt green dye, that exhibits greater fluorescence enhancement upon binding to doublestranded dna dsdna than sybr green i. Lightcycler 480 block kit 384well 04 643 631 001 96well 04 643 640 001 generic reagents lightcycler 480 sybr green i master 1 kit 500 reactions 04 707 516 001 lightcycler 480 probes master 1 kit 500 reactions 04 707 494 001 lightcycler 480 genotyping master 1 kit 384 reactions 04 707 524 001 other reagents. Quantitative analysis of copy number variants based on.
Oligonucleotide primers are dissolved in sdh 2 o to a stock concentration of 10. Faststart dna master sybr green i, using the lightcycler. However, roche diagnostics gmbh reserves the right to make any changes necessary without notice as part of. Executed on roche lightcycler 480 realtime pcr system. Reactions were run in duplicate on the lightcycler 480 using 10fold dilutions of human leukocyte cdna 100 ng to 100 pg and a quantitect primer assay for myc a protooncogene. In twostep rtpcr, the reverse transcription of rna into cdna is separated from the other reaction steps and is performed outside the. Despite this, the sensifast kit performed significantly better, leasing to earlier amplification from comparable genomic dna samples see picture for example. Lightcycler 480 instrument ii roche applied science. Lightcycler 480 sybr green i master lightcycler 480 sybr green i master. I am using life technologies sybr green master mix abi on a roche real time pcr machine lc96. The system features the lightcycler 480 instrument, a versatile, platebased realtime pcr device that supports mono or multicolor applications.
Lightcycler 96 realtime pcr system super capabilities are. If taking different reaction volumes, scale all components proportionally. Across an 8log dilution series of across an 8log dilution series of plasmid, fast sybr green master mix demonstrates superior dynamic range and linearity compared to competitor r. Roche applied science 4 application lightcycler 480 sybr green i master is designed for research studies. The use of maxima sybr green qpcr master mixes in realtime pcr ensures reproducible, sensitive and specific quantification of genomic, plasmid, viral, and cdna templates. Hot start protocols used with the lightcycler 480 sybr green i master have been shown to significantly. Lightcycler faststart dna masterplus sybr green i version october 2007 easytouse hot start reaction mix for pcr using the lightcycler carousel based system cat. Taqman fast advanced master mix on the roche lightcycler 480. Lightcycler 480 sybr green i master roche life science.
Thermo scientific maxima sybr green qpcr master mix 2x. Kapa sybr fast kit for lightcycler 480 sigmaaldrich. However, roche diagnostics gmbh reserves the right to make any changes necessary without notice as part of ongoing product development. Lightcycler 480 sybr green i master 5 x 1 ml 2x conc. The sybr green i dye contained in kapa sybr fast qpcr master mix 2x and roxfluorescein dyes depending on kit configuration are light sensitive. This sybr green pcr kit may be used to quantify target dna using either absolute or relative quantification. I am using the lightcycler 480 sybr green i master kit from roche, but it is brand new. These should be prepared fresh before each experiment. Kits include kapa sybr fast qpcr master mix 2x, which contains kapa sybr fast dna polymerase, reaction buffer, dntps, sybr green i dye, and mgcl 2 at a final concentration of 2. View the new lightcycler 480 biochemica article gene.
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