详细描述：
CAL Fluor®和Quasar®染料校准标准品可用于改进需要光谱校准的qPCR仪中的信号解卷积。它们使仪器能够存储每种染料的荧光图谱并控制荧光之间的相互串扰。串扰是指某种荧光信号泄漏到相邻滤光器中的一种现象。多重定量中多个靶点同时扩增但独立检测，这种串扰问题尤其需要注意。不同荧光报告基团的信号代表不同靶标分子的表达量，使用标准染料进行校准能够解决仪器的光谱重叠问题。
染料校准标准与荧光基团一起共价连接到oligo-dT（10-mer）上，以更好地模拟荧光探针的信号。使用染料校准作为参考，分析软件会分析在扩增期间每个荧光基团预期发出的荧光，然后减去从不适当的滤光器组件中的信号值。因此，使用T10-染料标准品进行校准后减少了串扰的幅度。
CAL Fluor®和Quasar®染料标准品跨越宽范围的发射波长，因此在实验设计中具有更大的灵活性。有关常用仪器光谱校准的说明请参阅以下链接：https://biosearch-cdn.azureedge.net/assetsv6/BTI_Spectral_Calibration_Instructions.pdf。在某种仪器上，用于多重定量的所有染料必须一起校准，因此了LGC Biosearch也提供FAM校准标准染料。若您需要定制合成某种校准染料，或者如果您对校准程序有任何疑问，请咨询info@genecompany.com。
CAL Fluor® and Quasar® dye calibration standards are available to improve signal deconvolution in real-time qPCR thermal cyclers that require spectral calibration. They enable the instrument to store the fluorescent profile of each dye and control for crosstalk. Crosstalk is the bleed-through of signal into adjacent filter-sets that are oriented to detect other fluorophores. This crosstalk is particularly consequential to a multiplexed assay, since multiple targets are amplified simultaneously but detected independently. Different fluorescent reporters are used to signal each target and so calibration with dye standards allows the instrument to resolve their overlapping spectra.
Dye calibration standards are formulated with the fluorophore covalently attached to an oligo-dT (10-mer), to better mimic the signal from a fluorescent probe. Using the dye calibration as a reference, the analysis software anticipates how much fluorescence to expect from each fluorophore during amplification, and will subtract out signal from inappropriate filter-sets. Calibration with the T10-dye standards therefore reduces the magnitude of crosstalk.
CAL Fluor and Quasar dye standards span a wide range of emission wavelengths for maximum flexibility in experimental design. Instructions on spectral calibration are linked below for a number of common instruments. On certain instruments, all dyes that will be multiplexed together must be calibrated together, and so a FAM standard is available as well. Please inquire with info@genecompany.com to discuss the custom synthesis of any dye not currently available as a calibration standard, or if you have any questions on the calibration procedure.
