Is anyone using life technologies sybr green master mix on. I am using the lightcycler 480 sybr green i master kit from roche, but it is brand new. It contains faststart taq dna polymerase and dna doublestrand. Lightcycler 480 sybr green i master lightcycler 480 probes master. Manual and the pack insert for lightcycler dna master sybr green i. The sybr green i dye contained in the kapa sybr fast qpcr master mix 2x and roxfluorescein dyes depending on kit configuration are light sensitive. I have a problem with the analysis in lightcycler 480. Lightcycler 480 realtime pcr system gene quantification. Oligonucleotide primers are dissolved in sdh 2 o to a stock concentration of 10. Is anyone using life technologies sybr green master mix on a. A rationale for atypical endoflagella article snippet. Ssoadvanced universal sybr green supermix is a 2x concentrated, readytouse reaction master mix optimized for dyebased realtime pcr on any realtime pcr instrument roxindependent and roxdependent.
Lightcycler 480 software release 1 5 0 sp4 roche bioz. Maxima sybr green qpcr master mix, rox solution privided. Start the lightcycler 480 software by doubleclicking the lightcycler480 icon. Powerup sybr green master mix ssoadvanced universal sybr green supermix biorad ssofast evagreen supermix biorad lightcycler 480 sybr green i master mix roche perfecta sybr green supermix quantabio quantitect sybr green pcr kit qiagen 10 1 0. Lightcycler 480 sybr green i master mix bioz ratings. Easeofuse, convenience, speed, reduced error result consistency, stability and tolerance of targets of varying secondary structure. Kapa sybr fast onestep qrtpcr master mix 2x kit kapa sybr. The complete pcr mix that is lightcycler 480 sybr green master. 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.
The software is compatible with the regulations for electronic records subpart b. Before preparing qpcr reactions, thoroughly mix the kapa probe fast bio rad icycler qpcr master mix 2x, template dna, primers and probes. Sign, fax and printable from pc, ipad, tablet or mobile with pdffiller. Transcript levels of nrf1, klf15, sp1 and hpcft were monitored by realtime rtpcr with a lightcycler 480 probes master kit, or with a lightcycler 480 sybr green i master kit. Lightcycler faststart dna master sybr green i readytouse hot start reaction mix for pcr using the lightcycler1 instrument cat. I am having a lot of issues while standardizing my reaction conditions. Since the mix is provided as an easytouse allinone master reagent, reaction setup only requires. Read the msds and follow the handling instructions. Lightcycler 480 software release 1 5 0 sp3 roche bioz. Benefit from cuttingedge thermal block and optical technology that leads to excellent welltowell homogeneity and maximized detection. The lightcycler 480 sybr green i master is a onecomponent hot start reaction mix for pcr. Qpcr protocol with sybr green for lightcycler 480 clinisciences.
It contains faststart taq dna polymerase and dna double strand. The pfaffl model and the relative expression software tool restmcs. Lightcycler faststart dna master sybr green i y version. Building on over 20 years of innovation and product excellence in qpcr, our powerup sybr green master mix is designed for exceptional performance for your most challenging realtime pcr applications. Powerup sybr green master mix thermo fisher scientific es. Brilliant iii ultrafast sybr green qrtpcr master mix quick. The kit is ideally suited for hot start pcr assays for gene detection and quantification. The lightcycler 480 software uses unique roche algorithms for highly accurate and robust automated data analysis. Opatreni pri pozaru maji odpovidat okolnim podminkam. 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. Quickly start experiments from readytouse macros or templates for all applications. Highthroughput meltingcurve analysis of samples containing different variants of the polymorphic mammalian mdr1 gene.
Transfer the multiwell plate into the plate holder of the lightcycler. Minimize exposure of maxima sybr green qpcr master mix 2x, no rox and rox solution. Typical sybr green reaction protocol reaction setup reagent final concentration volume l one reaction water 1. Wear appropriate protective eyewear, clothing, and gloves. Roche applied science lightcycler 480 realtime pcr system. Applied biosystems powerup sybr green master mix is a preformulated, optimized universal 2x master mix for realtime pcr. It contains faststart taq dna polymerase and dna doublestrandspecific sybr green i dye for pcr product detection and characterization. Lightcycler 480 sybr green i master roche life science.
Lightcycler 480 sybr green i master mix thermo fisher bioz. Lightcycler 480 sybr green i master 2x concentrated 04 707 516 001 04 887 352 001. The fast sybr green master mix protocol pn 4385372 provides detailed realtime pcr and rtpcr procedures and ordering information for the fast sybr green master mix. It contains antibodymediated hotstart sso7d fusion polymerase, dntps, mgcl2, sybr green i dye, enhancers, stabilizers, and a blend of.
Contents vial bottle cap label function catalog number content 1 green lightcycler 480 sybr green i master, 2x conc. For optimal results, recommended primer concentrations should. Readytouse hot start pcr mix contains faststart taq dna polymerase, reaction buffer. Furthermore, the lightcycler 480 software, version 1. Lightcycler 480 sybr green i master 10 x 5 ml 2x conc. The transcript levels were normalized to that for gapdh andor. In twostep rtpcr, the reverse transcription of rna into cdna is separated from the other reaction steps and is performed outside the. Kapa sybr fast qpcr master mix 2x is stable through 30 freezethaw cycles. When used with the lightcycler 480 system, this kit is ideally suited for hotstart pcr applications.
Lightcycler carouselbased instruments and lightcycler 480 instrument. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity. Readjust the threshold value for analysis of every run. The lightcycler480 sybr green i master provides reagents including sybr green i dye, for amplification and detection of dna using the lightcycler480 instrument. May 2017 easytouse reaction mix for onestep rtqpcr using the lightcycler 480 or the lightcycler 96 realtime pcr systems cat. Application lightcycler 480 sybr green i master is designed for research studies. Protect the lightcycler faststart reaction mix sybr green i. Lightcycler faststart dna masterplus sybr green i version october 2007 easytouse hot start reaction mix for pcr using the lightcycler carousel based system cat. I am using life technologies sybr green master mix abi on a roche real time pcr machine lc96. Enter username and password when the login dialog box appears and click the button to proceed with the login. 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.
Exceptional specificity, broad instrument capability. Powerup sybr green master mix thermo fisher scientific ae. Lightcycler 480 sybr green i master 5 x 1 ml 2x conc. The lightcycler evoscript rna sybr green i master is designed for sensitive, highspecificity, highprecision onestep rtqpcr especially for gene expression assays. Lightcycler 480 sybr green i master version june 2005 easytouse hotstart reaction mix for pcr using the lightcycler 480 system cat. Roche diagnosticslightcycler 480 sybr green i m fisher. Sybr green master lightcycler 480 sybr green i master 04 707 516 001 5 ml 5 x 20 l 2x concentrated master mix for 250 20 l reactions high resolution melting lightcycler 480 high resolution melting master 04 909 631 001 5 ml 5 x 1 ml 2x concentrated 25 ml mgcl2 stock solution for 500 20 l reactions hybridization probes.
Sybr green i is the most frequently used dsdnaspecific dye in qpcr. Lightcycler 480 instrument absolute quantification quick. Roche lightcycler 480 sybr green i master reaction mix lightcycler 480 sybr green i master reaction mix, supplied by roche, used in various techniques. Lightcycler 480 sybr green i master fill online, printable, fillable.
Lightcycler evoscript rna sybr green i master y version. Load the multiwell plate into the the lightcycler 480 instrument. Lightcycler 480 sybr green i master mix roche diagnostics. It is an asymmetric cyanine dye that largely binds to the minor groove of dsdna, independent of the nucleotide sequence. Powerup sybr green master mix thermo fisher scientific. Contains faststart taq dna polymerase, reaction buffer, dntp mix with dutp instead of dttp, sybr green i dye, and mgcl 2. Allow the fast sybr green master mix to thaw completely. All sybr green i assays have to be verified by a melting curve analysis for side products other than the specific amplicon. Brilliant iii ultrafast sybr green qpcr master mix quick.
Amplifications were carried out on a lightcycler 480 ii instrument using lightcycler 480 software v. January 2017 easytouse hot start reaction mix for pcr using sybr green i with the lightcycler carouselbased system cat. Introducing the lightcycler 480 system the next generation. Turn on the computer linked to the instrument and log on to window xp. Enter sample information for your experiment as needed. Sybr green i binds all doublestranded dna molecules, emitting a fluorescent signal on binding. Prvni pomoc musi byt zahajena za ucasti specialisty oboru pracovniho lekarstvi.
Enter the username and assigned password to log in to the. Genomic dna up to 500 ng and plasmid dna up to 10 ng can be used in qpcr with to light during handling to avoid lo ss of fluorescent signal intensity. It can be excited with blue light with a wavelength of 480 nm, and its emission spectrum is comparable to that of fluorescein with a maximum at 520 nm. Green supermix biorad lightcycler 480 sybr green i master mix roche perfecta sybr green supermix quantabio quantitect sybr green pcr kit qiagen in an evaluation of 24 different primer sets used with powerup sybr green master mix, a single melt curve was obtained in 100% of reactions without the need for primer optimization or redesign. Roche diagnostics lightcycler 480 sybr green i m fisher sci. Realtime pcr was performed with the same amount of cdna in lightcycler 480 multiwell plate 384 wells using roche lightcycler 480 sybr green i master or roche lightcycler 480 probe master roche diagnostics gmbh, mannheim, germany. Product bulletin powerup sybr green master mix p owerup sybr. Product bulletin powerup sybr green master mix powerup sybr. Product name reagents catalog number pack sizedescription lightcycler 480 sybr green i master 04 707 516 001 5 ml 5 x 1 ml lightcycler 480 probes master 04 707 494 001 5 ml 5 x 1 ml lightcycler 480 genotyping master 04 707 524 001 master mix, 5 x concentration, 4 x 384 l, readyto. 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. Fill lightcycler 480 sybr green i master, edit online. Doubleclick the lightcycler 480 icon on the desktop. Brilliant iii ultrafast sybr green qrtpcr master mix.
Program the lightcycler 480 instrument before preparing the reaction mixes. For in vitro lightcycler faststart dna master sybr green i. Magnesium chloride kapa sybr fast qpcr master mix 2x contains an optimized. The realtime pcr amplifications were performed using the lightcycler 480 sybr green i master roche, 50 nm gene specific primers additional file. In combination with the lightcycler 480 system and suitable pcr primers, this kit allows very sensitive detection and quantification of defined dna sequences. Contents vialbottle cap label function description content 1 red lightcycler rna master sybr green i, lc rna master sybr green l, 2. Lightcycler 480 sybr green i master roche diagnostics.
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