Sybr green i master mixture of 100 mm trishcl, ph 8. In our lab, we use power sybr green pcr master mix from applied biosystems ref 4367659. Introduction to taqman and sybr green chemistries for real. It is supplied in a convenient 2x concentration premix to perform realtime. Sybr green realtime pcr master mix, quantitative pcr master. The mix is provided as an easytouse master reagent. This protocol is for the comparison of powerup sybr green master mix to one other master mix using 20 l reactions in. Each reaction had a total volume of 25 l, including 12. Sybr green pcr master mix and rtpcr reagents protocol. The sybr green i dye contained in kapa sybr fast qpcr master mix 2x and roxfluorescein dyes depending on kit configuration are light sensitive. Power sybr green pcr master mix contains a blend of dttpdutp that is compatible with amperase ung, to minimize carryover contamination includes a proprietary version of rox dye, an internal passive reference, to normalize nonpcrrelated fluorescence fluctuations to minimize welltowell variability that result from a variety of. Brilliant iii ultrafast sybr green qrtpcr master mix. It contains faststart taq dna polymerase and doublestranded dna specific sybr green i dye for product detection and characterization.
With the bioline sensifast mix, our targets regularly appear at lower cp values and all primers tested work more reliably and consistently see attached table for two examples of primer efficiencies determined with the lightcycler and the sensifast kits. The system includes a proprietary dsdnabinding dye, carboxyx. Smobio sybr green realtime pcr master mix, sybr exceltaq 2x qpcr master mix sybr, rox. Ssoadvanced universal sybr green supermix is an exclusive highperformance realtime pcr reagent based on biorads patented sso7d fusion protein polymerase technology and advanced buffer formulation. Faststart essential dna green master roche life science. Fyi, the amplification efficiency for all primers have been validated using the commercial sybr green master mix, and were running our samples on a cfx96 realtime detection system biorad. Additional equipment and reagents required additional reagents and equipment required to perform reactions with the lightcycler 480 sybr green i master. During each phase of dna synthesis, the sybr green i dye, which is included in the reaction mix, binds to the amplified pcr products. Brilliant ii sybr green qrt pcr master mix 1step, part number 600825 section 2.
Sybr green i is a commonly used fluorescent dye that binds doublestranded dna molecules by intercalating between the dna bases. In our own experiments, we found that powertrack sybr green master mix outperformed other master mixes when comparing specificity, pcr efficiency, dynamic range, and precision over multiple targets. Robust performance of powerup sybr green master mix over a broad dynamic range. For longer storage, this reagent should be kept at 20c and protected from light. Brilliant ii sybr green qrtpcr master mix with rox. The brilliantii master mix is also ideal for quantification of cdna in a 2step qrtpcr reaction when combined with the affinityscript qpcr cdna synthesis kit. Protocol powerup sybr green master mix guidelines for. White paper sybr green assays design and optimization of sybr. Quantitect sybr green pcr kits overcome the need for optimization of reaction conditions, which can be tedious and timeconsuming. Can anyone recommend a protocol for sybr green master mix. Sybr green realtime pcr master mixes thermo fisher. It contains faststart taq dna polymerase and doublestranded dna specific sybr.
Follow the protocol in the handbook to get fast and reliable results on any realtime. Applied biosystems sybr green pcr master mix combines sybr green i dye, amplitaq gold dna polymerase, dntps with dutp, passive reference. Applied biosystems sybr green master mixes are designed for quantitative realtime pcr using a set of two pcr primers that flank the target region. Power sybr green pcr master mix and power sybr green rt pcr reagents kit user guide 9 1 product information purpose of the kit the power sybr green pcr master mix is a convenient premix of the components except primers, template, and water necessary to perform realtime pcr using sybr green i dye with enhanced sensitivity and specificity. The master mix contains all required components, except rox. It is supplied in a convenient 2x concentration pre mix to perform realtime pcr using sybr green dye. Power sybr green pcr master mix contains a blend of dttpdutp that is compatible with amperase ung, to minimize carryover contamination includes a proprietary version of rox dye. Pcr master mix certificate of analysis 9pim750 promega. Powerup sybr green master mix degrades any previously amplified pcr products and helps prevent contamination of subsequent qpcr reactions.
Guaranteed minimum shelf life is 60 days exact expiry date printed. Utilization of a heatlabile ung enzyme allows for effective contamination control figure 3. Kapa sybr fast qpcr master mix 2x is stable through 30 freezethaw cycles. Applied biosystems sybr green pcr master mix combines sybr green i dye, amplitaq gold dna polymerase, dntps with dutp, passive reference 1, and optimized buffer in the convenience of a single vial. Primers should be designed according to the following guidelines. For all chemicals in bold red type below, read the msds and follow the handling instructions. We have compared the mix to the roche lighcycler 480 sybr green i master mix. Sybr green pcr master mix applied biosystems, cat log. These are available online in convenient and compact pdf. Guaranteed minimum shelf life is 60 days exact expiry date printed on product and cofa. The brilliant iii sybr green ultrafast qrtpcr master mix was developed for the abi steponeplus and biorad cfx96 realtime pcr instruments and other fastcycling systems. Quantitative rtpcr protocol sybr green i schnable lab.
This product is a taq dna polymerasebased 2 x master mix for realtime pcr, which contains all components, except for the primer. Upon binding dsdna, the fluorescence of sybr green i increases by approximately fold, making it ideal for detecting the accumulation of dsdna. Direct detection of pcr product is monitored by measuring the increase in. Pcr master mix is a premixed, readytouse solution containing taq dna polymerase, dntps, mgcl2 and reaction buffers at optimal concentrations for efficient. Gotaq qpcr master mix is a simpletouse, stabilized 2x formulation that includes. For safety and biohazard guidelines, refer to the safety. Simply add primers and dna template to the readytouse pcr master mix, and start the reaction see flowchart twostep rt pcr. Read the safety data sheets sdss and follow the handling instructions. The use of sybr green i for detection in realtime pcr has a cost advantage, but data are susceptible to misinterpretation because of the presence of pcr artifacts. The applied biosystems powerup sybr green master mix is formulated to provide superior specificity and sensitivity. It contains antibodymediated hotstart itaq dna polymerase, dntps, mgcl 2, sybr green i dye, enhancers, stabilizers, and a blend of passive reference dyes including rox and.
For the best quantitation results, use the following. Power sybr green pcr master mix thermo fisher scientific. Product bulletin powerup sybr green master mix powerup sybr. The master mix contains all required components, except rox reference dye and primers 50x rox reference dye is individually supplied with this kit. This reagent can be stored at 4c for 2 months and protected from light. Do qrtpcr and test the selected primers 1 qrtpcr set up. Power sybr green pcr master mix and power sybr green rt. 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. A protocol for a stabilized 2x master mix that includes all components for quantitative pcr except sample dna, primers and water. Applied biosystems power sybr green pcr master mix 1 x 5ml. Hot start protocols used with the lightcycler 480 sybr green i master have been shown to significantly.
The faststart essential dna green master is a onecomponent hot start reaction mix for qualitative and quantitative pcr. Executed on biorad cfx96 realtime pcr detection system. Gotaq green master mix contains two dyes blue and yellow that allow monitoring of progress during electrophoresis. In all the sets, cdna was amplified by a realtime pcr using power sybr green pcr master mix kit abi, usa in onestep abi instrument abi, usa. Rtrnase block enzyme mixturewarning hazard statements. Sybr green pcr master mix and sybr green rt pcr reagents kit user guide 9 1 product information purpose of the kit the sybr green pcr master mix is a convenient premix of the components except primers, template and water necessary to perform realtime pcr using sybr green i dye. Sybr green realtime pcr master mix, sybr green quantitative pcr master mix.
Brilliant iii ultrafast sybr green qrtpcr master mix 3 introduction quantitative reverse transcription pcr qrtpcr is a powerful tool for gene expression analysis. Faster qrtpcr master mixes, brilliant iii sybr master mix. Ssoadvanced universal sybr green supermix is an exclusive highperformance realtime pcr reagent based on biorads patented sso7d fusion protein polymerase technology and advanced buffer. To maintain optimal performance of the quantitect sybr green rt pcr kit for x 50 l reactions, we recommend storing the 25 ml master mix as appropriately sized aliquots in sterile, polypropylene tubes. To circumvent this problem, we carefully optimize the reaction conditions for each primer pair to minimize nonspecific products or primer dimers and to maximize yield of the desired amplicon. Rt mix must always be stored at 20c to guarantee performance. Lightcycler 480 sybr green i master roche life science. It is used in quantitative pcr because the fluorescence can be measured at the end of each amplification cycle to determine, relatively or absolutely, how much dna has been amplified. In our own experiments, we found that powerup sybr green master mix outperformed other master mixes when comparing specificity, pcr efficiency, dynamic range, and precision over multiple targets. Sybr green pcr master mix and sybr green rtpcr reagents kit user guide 9 1 product information purpose of the kit the sybr green pcr master mix is a convenient premix of the components except primers, template and water necessary to perform realtime pcr using sybr green i dye.
The purchase of this product from applied biosystems includes a limited, nontransferable immunity from suit under the foregoing patent claims for using only this. Brilliant iii ultrafast sybr green qpcr master mix quick reference guide for the abi 7500 fast realtime pcr system contains information on the brilliant iii ultrafast sybr green qrtpcr master mix with the materials provided, materials needed, pcr protocols, storage conditions, and msds. Instruction manual sybr green realtime pcr master mix plus 0803 f0925k sybr green realtime pcr master mix plus qpk212 1mlx5 store at 20c, protected from light contents 1. Gotaq qpcr master mixa,b is a reagent system for quantitative pcr qpcr. Ssoadvanced universal sybr green supermix life science. For quantitative, realtime pcr and twostep rtpcr using sybr green i pdf 142kb english format file size. Powerup sybr green master mix universal 2x master mix for realtime pcr workflows catalog numbers a25741, a25742, a25743, a25776, a25777, a25778, a25779, a25780, a25918 pub. Quantitative rtpcr protocol sybr green i 4 quantitative realtime pcr qrtpcr 1. In rna quantitation assays, you use the sybr green pcr master mix in the second step of a twostep reversetranscription polymerase chain reaction rt pcr. Sufficient reagents provided for 40 reactions based on a 50 l reaction volume. Evaluation of a homemade sybr green i reaction mixture for. Thunderbird sybr qpcr mix is a taq dna polymerasebased highly efficient 2x master mix for realtime pcr using sybr green i. An added benefit of an evagreenbased master mix is that you can analyze your pcr product by gel electrophoresis without the need to add another dnabinding dye to either your loading buffer or gel. Sybr green dye duallock dna polymerase, with a proprietary combination of.
Do two reactions for each pair of primers by using cdna and h2o as templates separately. The master mixes contain buffer, dntps, thermostable hotstart dna polymerase, and sybr green dyeeverything needed for realtime pcr. In our own experiments, we found that powerup sybr green master mix outperformed other master mixes when comparing specificity, pcr efficiency, dynamic range, and. The purchase of this product from applied biosystems includes a limited, non. Powerup sybr green master mix, 10pack 10 x 1 ml 1,000 reactions a25918 powerup sybr green master mix, 5pack 5 x 5 ml 2,500 reactions a25777 powerup sybr green master mix, bulk pack 1 x 50 ml 5,000 reactions a25743 powerup sybr green master mix, 10pack 10 x 5 ml 5,000 reactions a25778 to learn more about taqman assays. It is supplied in a convenient 2x concentration premix to perform realtime pcr using sybr green dye. The complete pcr mix that is lightcycler 480 sybr green master supplemented with primers and template is stable for up to 24 h at room temperature. Sybr green pcr master mix and sybr green rtpcr reagents. Wear appropriate protective eyewear, clothing, and gloves. Powerup sybr green master mix, 10pack 10 x 1 ml 1,000 reactions a25918 powerup sybr green master mix, 5pack 5 x 5 ml 2,500 reactions a25777 powerup sybr green master mix, bulk pack 1. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity. Sybr green master mix to applied biosystems powertrack sybr green master mix using this simple protocol. Sybr green realtime pcr master mix, quantitative pcr master mix. The evagreen dye in the master mix can act as a dna prestain, permitting direct visualization of dna bands following electrophoresis.
Premixed components stored at 2 to 8c significantly reduce assay setup time. Performing realtime pcr assays quick reference card for safety and biohazard guidelines, refer to the safety section in the power sybr green pcr master mix and rtpcr protocol pn 4367218. Use of the power sybr green pcr master mix is covered by u. Will the qpcr work with an expired sybr green master mix. White paper sybr green assays design and optimization of. The neb luna universal qpcr master mix is an optimized 2x reaction mix for realtime qpcr detection and quantitation of target dna sequences using the sybr fam channel of most realtime qpcr instruments. Sybr green qpcr kits radiant sybr green qpcr kits are designed for the majority of applications for the fast, sensitive detection of cdna, genomic dna and viral targets. Applied biosystems sybr green pcr master mix combines sybr green i dye, amplitaq gold dna polymerase, dntps with dutp, passive reference 1, and optimized buffer in the convenience of a. Sybr green realtime pcr master mix 1 ml sybr green realtime pcr master mix 1 ml x 5 notes. Power sybr green pcr master mix and rtpcr protocol pn.
Sybr green pcr master mix thermo fisher scientific. Gotaq green master mix is a premixed readytouse solution containing bacterially derived taq dna polymerase, dntps, mgcl 2 and reaction buffers at optimal concentrations for efficient amplification of dna templates by pcr. The reproducibility and sensitivity of gene expression analyses by quantitative realtime pcr was increased by the. Sybr green pcr master mix and rtpcr reagents protocol pn. The sybr green pcr master mix is designed for use with the abi prism 7700 sequence detection system sds, the abi prism 7900ht sds, the abi prism 7000 sds, or the geneamp 5700 sds. Smobio sybr green realtime pcr master mix, sybr exceltaq 2x qpcr master mix sybr, rox, tq1110 is a readytouse reagent with all the essential components for quantitative realtime pcr qpcr except primers and template. Simply add primers and dna template to the readytouse pcr master.
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