Dye or Probe Based QPCR

DYE - OR PROBE- BASED qPCR Dye- or Probe Based qPCR/RT-qPCR How to Choose D Dye-Based qPCR/RT-qPCR GoTaq® qPCR and RT-qPCR Systems are ready-to-use 2X master mixes optimized for qPCR using a dsDNA binding dye. The systems contain BRYT Green® Dye, which has a greater fluorescence enhancement upon binding to dsDNA than does SYBR® Green I. The 1-Step and 2-Step RT-qPCR Master Mixes contain GoScript™ Reverse Transcriptase for efficient first-strand cDNA synthesis. In addition, the Master Mixes are 2X, and contain all required components; just add primers and template. GoTaq® qPCR Master Mix provides robust real-time PCR with earlier quantification cycle values and broad range detection for increased reliability, reproducibility and sensitivity. A. Delta Rn vs. Cycle GoTaq® Fluorescence Higher 0.01ng 0.1ng 100ng 10ng 1ng Vendor L Earlier Cq Values Cycle Number 0.01ng 0.1ng 1ng 10ng 100ng qPCR Master Mix B. 10 15 20 25 30 35 0 5 Promega Vendor L ye-based quantitative PCR (qPCR) uses a double-stranded DNA (dsDNA) binding dye such as BRYT Green® to detect the accumulation of amplification products after each reaction cycle. Probe-based qPCR monitors the fluorescent signal that is released by a fluorescently-labeled probe (e.g., TaqMan®) at the end of each reaction cycle. Here we help you choose the type of assay that best suits your studies. 100ng 17.57 20.26 10ng 20.75 23.23 1ng 24.19 26.43 0.1ng 27.65 29.87 0.01ng 30.89 32.60 Figure 1. A series of five tenfold dilutions of human gDNA was amplified with GoTaq® qPCR Master Mix or Vendor L’s equivalent master mix, to examine GAPDH expression. GoTaq® qPCR Master Mix shows a higher fluorescent signal (Panel A) and earlier Cq values (Panel B) for all sample input amounts. All no-template control reaction wells were blank. enews | September 2015 Delta Rn 8277TD Cq Cover

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