BioTechniques · 2018 · 82 citations · 9 references
Life SciencesBiotechnologyMolecular BiologyGene ExpressionGenomicsSynthetic BiologyNucleic Acid Amplification TestingOligonucleotide ChemistryPolymerase Chain ReactionPolymerase Chain Reaction (Pcr) AssaysProbe-based AssaysQuantitative Pcr DetectionReal-time Polymerase Chain Reaction (Pcr)Standard Curve ConstructionStandard DevelopmentTechnical Standard Development
Real-time PCR (qPCR) is widely used in the life sciences. For quantifying DNA, a standard curve is required. Common methods for standard development are time consuming, costly, necessitate a specific skill set, and pose a contamination risk. Using a targeted synthetic oligonucleotide, such as a gBlocks® Gene Fragment, overcomes these drawbacks and provides researchers an accurate and quick solution to standard development. Here, we demonstrate that using a gBlocks fragment as a standard provides comparable sensitivity, reliability, and assay performance to a purified amplicon standard.
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