QuantiFast Probe Assays
For qPCR and qRT-PCR gene expression analysis using predesigned, bioinformatically-verified assays together with QuantiFast Kits
QuantiFast Probe Assays are predesigned assays based on dual-labeled hydrolysis probe detection for gene expression analysis. Each assay for a specific gene is supplied as a lyophilized mix of forward and reverse primers and probe that can easily be reconstituted to obtain a 20x assay solution. Used in combination with QuantiFast Kits, QuantiFast Probe Assays enable rapid real-time RT-PCR quantification of cDNA or RNA targets in a singleplex format (FAM reporter dye) or duplex format (FAM and MAX reporter dyes). QuantiFast Probe Assays are available for every human, mouse, or rat gene. They are designed to enable amplification and detection of all targets, including ones that are less than 100 bp in size, which makes them highly suited for use with degraded starting material such as formalin-fixed, paraffin-embedded (FFPE) analytes. For convenience, the master mix can be stored at 2–8°C.
QuantiFast Probe Assays are designed for gene expression analysis and, in addition, are highly suited for FFPE analytes (see figure "Highly efficient real-time RT-PCR detection using RNA from an FFPE sample" and "Highly efficient real-time RT-PCR detection of degraded RNA").
QuantiFast Probe Assays are predesigned, genomewide assays for human, mouse, or rat genes. They are designed using a proprietary algorithm to enable amplification and detection of any RNA and cDNA targets. In addition, they can detect fragments of less than 100 bp in size with high efficiency and reliability. Each QuantiFast Probe Assay consists of a premixed primer pair and a dual-labeled probe. Two dye labels are available: FAM (for all genes) and MAX (for selected reference genes). QuantiFast Probe Assays can be easily ordered at the GeneGlobe Web portal.
For single or duplex one-step RT-PCR, QuantiFast Probe Assays are used in combination with the QuantiFast Probe RT-PCR Plus Kit for the detection of RNA. After addition of the master mix, reverse transcription and real-time PCR take place in the same tube. This kit offers an integrated genomic DNA removal step that effectively eliminates genomic DNA contamination to avoid false-positive signals. Removal of genomic DNA is particularly important in samples containing highly degraded RNA (e.g., RNA from FFPE tissue samples), where the presence of genomic DNA frequently results in false positive RT-PCR signals.
For single or duplex two-step RT-PCR, QuantiFast Probe Assays are delivered in combination with the QuantiFast Probe PCR Kit or the QuantiFast Multiplex PCR Kit, respectively. QuantiFast Probe Assays, used together with QuantiFast Kits, provide reliable and highly sensitive detection of cDNA targets and eliminate tedious design and optimization steps.
For cDNA synthesis, the QuantiTect Reverse Transcription Kit is recommended. The QuantiTect Reverse Transcription Kit provides an easy and fast cDNA synthesis procedure and includes a genomic DNA elimination step to avoid false-positive signals.
Each QuantiFast Probe Assay is supplied as a lyophilized mix of forward and reverse primers and a dual-labeled probe. QuantiFast Probe Assays should be reconstituted in TE, pH 8.0 (provided) to give a 20x mix, which is then added to the master mix supplied with a QuantiFast Kit. The reaction mix is aliquoted into PCR tubes, wells, or Rotor-Discs, and the individual template samples are then added.
For guaranteed results in gene expression analysis, QuantiFast Probe Assays are provided with the following kits shown in the table.
For both singleplex and duplex two-step qRT-PCR assays, cDNA can be prepared using the QuantiTect Reverse Transcription Kit, which provides cDNA synthesis with integrated removal of genomic DNA contamination.
Each QuantiFast Probe Assay available for a particular gene is given a number that appears in the product name after the gene symbol. For more information, please see the GeneGlobe FAQs.
QuantiFast Probe Assays are highly suited for applications such as gene expression analysis of targets less than 100 bp in size, which makes them highly suited for use with degraded starting material, including FFPE analytes.
For guaranteed results in gene expression analysis, QuantiFast Probe Assays are provided with the following kits: