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. At a point where the fluorescence signal is confidently detected over the background fluorescence, a quantification cycle, or Cq value, can be determined. Cq 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.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. 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). It also features dUTP for carryover prevention and a non-fluorescent, visible dye to monitor reaction setup. This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection.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. 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.Figure 1: The Luna Universal qPCR Master Mix offers exceptionalsensitivity, reproducibility and qPCR performanceqPCR targeting human GAPDH was performed using the Luna Universal qPCR Master Mix over a 6-log range of input template concentrations (20 ng – 0.2 pg Jurkat-derived cDNA) with 8 replicates at each concentration. cDNA was generated from Jurkat total RNA using the NEB Protoscript® II First Strand cDNA Synthesis Kit (NEB #E6560).Figure 2: The Luna Universal qPCR Master Mix provides sensitive and accuratedetection and quantitation across a wide variety of DNA sources The Luna Universal qPCR Master Mix is compatible with a broad range of genomic DNA sources. qPCR targets were quantitated with 50 ng – 0.5 pg genomic DNA as input using an ABI 7500 Fast real-time instrument. Genomic DNA was purified by typical column-based methods. In these examples, strong performance can be observed in the amplification of ACTB (encoding β-actin) from Mouse kidney genomic DNA, psbB (Photosystem II CP47 reaction center protein PsbB) from Tobacco, and RDN18 (18S ribosomal RNA) from Yeast.Figure 3: Extensive performance evaluation of commercially available dye-based qPCR reagents demonstrates the robustness and specificity of LunaqPCR reagents from NEB and other manufacturers were tested across 16–18 qPCR targets varyingin abundance, length and %GC, using either Jurkat genomic DNA or Jurkat-derived cDNA as input (10 genomic DNA targets and 8 cDNA targets on Bio-Rad real-time instrument, 9 genomic and 7 cDNA targets on ABI instrument). For each testing condition, data was collected by 2 users and according to manufacturer’s specifications. Results were evaluated for efficiency, low input detection and lack of non-template amplification (where ΔCq = average Cq of non-template control – average Cq of lowest input). In addition, consistency, reproducibility and overall curve quality were assessed (Quality Score). Bar graph indicates % of targets that met acceptable performance criteria (indicated by green box on dot plot and Quality Score > 3). Results for NEB and other major manufacturers are shown: Bio-Rad, SsoAdvanced™ Universal SYBR® Green Supermix; Roche, FastStart™ SYBR Green Master; QIAGEN, QuantiTect® SYBR Green PCR Kit; ABI, PowerUP™ SYBR Green Master Mix; Promega®, GoTaq® qPCR Master Mix. The Luna Universal qPCR Master Mix outperformed all other reagents tested. Learn more about our comprehensive qPCR/RT-qPCR testing and “dots in boxes” data visualizationFigure 4: The Luna Universal qPCR Master Mix contains an inert blue tracking dye to eliminate pipetting errors.
This product is related to the following categories:
Luna® qPCR & RT-qPCR Products,
PCR, qPCR & Amplification Technologies Products
This product can be used in the following applications:
购物车为空，快去购物吧！
4000-520-616
咨询顾问
特别 提示
