Genomic DNA or cDNA of interest can be quantitated with Luna qPCR, and existing as well as commercial qPCR assay primer sequences can be used.įigure 1: The Luna Universal qPCR Master Mix offers exceptional The master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification and quantitation of DNA except primers and DNA template. This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection. It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. It contains Hot Start Taq DNA Polymerase and has been formulated with a unique passive reference dye that is compatible across a variety of instrument platforms (including those that require a high or low ROX reference signal). The Luna Universal qPCR Master Mix is an optimized 2X reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR ®/FAM channel of most real-time qPCR instruments.
#Powerup sybr green master mix series#
C q values can be used to evaluate relative target abundance between two or more samples, or to calculate absolute target quantities in reference to an appropriate standard curve, derived from a series of known dilutions. At a point where the fluorescence signal is confidently detected over the background fluorescence, a quantification cycle, or C q value, can be determined. An internal reference dye, such as ROX, corrects well-to-well optical variations, and is used for fluorescent signal normalization.Dye-based quantitative PCR (qPCR) uses real-time fluorescence of a double-stranded DNA (dsDNA) binding dye, most commonly SYBR ® Green I, to measure DNA amplification during each cycle of a PCR. Many PCR machines require a passive reference dye. SYBR ® Green master mixes are available with other dyes including ROX. SYBR ® Green master mixes are 2x concentrated, and contain dNTPs, MgCl 2, and DNA polymerase. Nucleic acid amplification and expression profiling.SYBR ® Green master mix applications include: SYBR ® Green master mixes are compatible with any real-time PCR amplification protocol. SYBR ® Green Master Mix Applications and Uses The lack of a requirement for sequence- specific probes can result in shorter experimental preparation times. Most probe systems require different probes for different sequences. Not only is SYBR ® Green generally less expensive than using probes, it can be used with any combination of primers and template to monitor synthesis of all dsDNA sequences.
When the oligo primes a PCR reaction and is incorporated into a PCR product, the fluorescent molecule is no longer quenched. Generally, probes designed for PCR are oligos that have a fluorescent molecule on one end and a quencher on the other. One advantage of using a SYBR ® Green master mix is that it is cost effective, particularly when compared to probe-based detection systems. SYBR ® Green is not PCR inhibitory and provides accurate, reproducible results with high sensitivity over a broad dynamic range, and gives consistent results across different platforms. The quantity of dsDNA product in the reaction is proportional to the amount of fluorescence. As the PCR reaction proceeds, at each round of amplification SYBR ® Green dye binds to dsDNA as it polymerizes, resulting in an increase in the level of fluorescence at the end of each extension step. SYBR ® Green dye is a fluorescent double-stranded DNA (dsDNA)- binding dye that is used to track the progress of DNA amplification in real-time PCR experiments.
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