https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/dye-calibration-standards 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.

https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/molecular-beacons#?tab=product-info 分子信标是一类具有猝灭基团和荧光报道基团的双标记FRET探针。其与传统的双标记探针的区别在于分子信标在3'和5'末端掺入了一段短的（约5 bp）互补的茎序列。 这些互补序列杂交能够形成报告基团和猝灭基团在空间上紧靠在一起的“茎环”结构。这种替代构象使分子信标具有很强的分辨能力，能够识别仅具有单个核苷酸差异的不同目标序列。分子信标另一个不同于传统双标记探针之处在于它们不依赖于由Taq聚合酶的5'外切核酸酶活性引起探针水解产生荧光，因此，PCR后的溶解曲线是可能的。 就像标准双标记BHQ®探针一样，分子信标也进行多重定量。多重定量使用多个光谱分辨的荧光探针能够同时检测几个靶标。 LGC Biosearch分子信标得到了来自新泽西州纽瓦克的公共健康研究所许可进行销售。有关分子信标探针设计信息，请参阅公共健康研究所的分子信标网站。 Molecular Beacons are a type dual-labeled FRET probes incorporating a quencher and a fluorophore reporter molecule. They differ from traditional dual-labeled probes, due to the incorporation of a short (~5 b.p.) complementary stem sequence at the 3' and 5' ends. These complementary sequences hybridize to form a “stem-loop” structure which holds the reporter and quencher close together in space. This alternate conformation makes Molecular Beacons more discriminatory, allowing them to resolve different target sequences differing by a single nucleotide. Molecular Beacons also differ from traditional dual-labeled probes because they do not rely on probe hydrolysis from the 5' exonuclease activity of Taq polymerase to generate its fluorescence, therefore, a post-PCR melt curve is possible. Just like standard Dual-labeled BHQ® probes it is possible to multiplex assays using Molecular Beacons. Multiplexing allows for the detection of several targets simultaneously using multiple spectrally resolved fluorescent probes. Molecular Beacons are sold under license from the Public Health Research Institute, Newark, NJ. For information on designing Molecular Beacons please refer to The Public Health Research Institute's Molecular Beacons website.

https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhq-probes#?tab=product-info 双标记的BHQ探针是一种传统的线性FRET探针，其将荧光基团和淬灭基团共价连接到通常长度为20至30个碱基的寡核苷酸的5'和3'末端。荧光通过Taq聚合酶的5'外切核酸酶活性释放，Taq聚合酶在探针与其互补序列杂交时切割荧光染料。双标记BHQ探针是检测特异性靶序列的存在和定量的理想选择。 BHQ探针提供两种形式： 1）探针和引物在分开在单独管中形式； 2）用于基因表达和qPCR的ValuMix形式，即在单管中包含一个探针和两个引物的混合物。 ValumixPrimer的数量可以根据指定的探针量进行灵活地调整，如0.5 nmol，5 nmol或20 nmol。探针与引物的比率可以根据您的喜好在1：1至1：4.5的范围内。ValuMix是一个经济的选择，LGC Biosearch以经济实惠的价格提供0.5 nmol超小包装FAM-BHQ探针引物混合物。有关完整的选择范围，请参阅官网产品列表选项卡。 BHQ探针设计注意事项： 双标记探针作为一种末端标记探针，当序列长于30个碱基时会影响荧光淬灭效率。对于这种情况，请考虑BHQnova双猝灭探针（https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhqnova-probes）或将猝灭剂放在探针中间位置设计具有内部猝灭的探针（该形式当前仅提供FAM标记）。相反，短于20个碱基的序列可以被设计为小沟结合剂（MGB）探针，并且不可能在70℃的适当温度下结合。请考虑订购BHQplus®探针，或者联系基因有限公司技术人员咨询有关探针设计的任何问题。 注意： 1. 设计具有内部猝灭的探针（当前仅提供FAM） 2. 添加由T（...）注释的内部修饰表示胸腺嘧啶碱基连接到内部修饰。请注意，在添加此类内部修饰时，将在序列中引入另外的T碱基。 3. 具有不确定/降解碱基的序列如何设计探针，请参考https://www.biosearchtech.com/support/faqs/custom-oligonucleotides-modifications/what-are-wobbles 4. 为了确保寡核苷酸能被有效识别，当在序列输入框中输入不确定/降解碱基时请使用IUPAC代码，而非括号，如：（A / G），（A / G / C / T）等。 Dual-Labeled BHQ Probes are traditional, linear, FRET probes incorporating a fluorophore and quencher covalently attached to the 5' and 3' ends of an oligo typically 20 to 30 bases in length. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridization to its complementary sequence. Dual-Labeled BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences. BHQ probes are offer delivery of the probe and primers in separate individual tubes. We also offer ValuMix for gene expression and qPCR which delivers one probe and two primers in a single tube. ValumixPrimer quantities can be flexibly adjusted around a specified probe amount of either 0.5 nmol, 5 nmol, or 20 nmol delivered. Probe to primer ratios can range anywhere from 1:1 to 1:4.5 according to your preference. ValuMix is an economical option providing 0.5 nmol of FAM-BHQ probe mixed with primers for only $79. See the product listing tab for full range of options. Design Concerns for BHQ probes Please be aware that sequences longer than 30 bases are unlikely to have efficient quenching as an end-labeled probe. For such situations please consider positioning the quencher internally instead. Conversely, sequences shorter than 20 bases may have been designed as a Minor Groove Binder (MGB™) probe and are unlikely to bind at the proper temperature of 70 °C. Please consider ordering such a sequence as a BHQplus® probe instead, or else contact info@genecompany.com with any questions regarding probe design. NOTE: Building Probes with an Internal Quencher (Currently available with FAM only) Adding an internal modification annotated by T(...) indicates that a Thymidine base is attached to the internal modification. Please be aware that an additional T base will be introduced into the sequence when adding such an internal modification. Building Probes with Wobble/Degenerate Bases (What are Wobbles?) To ensure the most efficient processing of your oligo, use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc. Best DEAL: ValuProbe™ BHQ Probe - 10 nanomoles of FAM-BHQ Probe typically ships in 2 business days Instrument CALIBRATION: FAM, CAL Fluor and Quasar calibration dyes are available here for online purchase Learn MORE: Applying dual-labeled probes in Multiplexing qPCR, Gene Expression, and SNP Genotyping assays The difference between FRET and static quenching Assay DESIGN: Try RealTimeDesign™ Software, a FREE, qPCR assay design program. Design your Dual-Labeled BHQ probes and primers online today!

https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhq-probes#?tab=product-info 双标记的BHQ探针是一种传统的线性FRET探针，其将荧光基团和淬灭基团共价连接到通常长度为20至30个碱基的寡核苷酸的5'和3'末端。荧光通过Taq聚合酶的5'外切核酸酶活性释放，Taq聚合酶在探针与其互补序列杂交时切割荧光染料。双标记BHQ探针是检测特异性靶序列的存在和定量的理想选择。 BHQ探针提供两种形式： 1）探针和引物在分开在单独管中形式； 2）用于基因表达和qPCR的ValuMix形式，即在单管中包含一个探针和两个引物的混合物。 ValumixPrimer的数量可以根据指定的探针量进行灵活地调整，如0.5 nmol，5 nmol或20 nmol。探针与引物的比率可以根据您的喜好在1：1至1：4.5的范围内。ValuMix是一个经济的选择，LGC Biosearch以经济实惠的价格提供0.5 nmol超小包装FAM-BHQ探针引物混合物。有关完整的选择范围，请参阅官网产品列表选项卡。 BHQ探针设计注意事项： 双标记探针作为一种末端标记探针，当序列长于30个碱基时会影响荧光淬灭效率。对于这种情况，请考虑BHQnova双猝灭探针（https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhqnova-probes）或将猝灭剂放在探针中间位置设计具有内部猝灭的探针（该形式当前仅提供FAM标记）。相反，短于20个碱基的序列可以被设计为小沟结合剂（MGB）探针，并且不可能在70℃的适当温度下结合。请考虑订购BHQplus®探针，或者联系基因有限公司技术人员咨询有关探针设计的任何问题。 注意： 1. 设计具有内部猝灭的探针（当前仅提供FAM） 2. 添加由T（...）注释的内部修饰表示胸腺嘧啶碱基连接到内部修饰。请注意，在添加此类内部修饰时，将在序列中引入另外的T碱基。 3. 具有不确定/降解碱基的序列如何设计探针，请参考https://www.biosearchtech.com/support/faqs/custom-oligonucleotides-modifications/what-are-wobbles 4. 为了确保寡核苷酸能被有效识别，当在序列输入框中输入不确定/降解碱基时请使用IUPAC代码，而非括号，如：（A / G），（A / G / C / T）等。 Dual-Labeled BHQ Probes are traditional, linear, FRET probes incorporating a fluorophore and quencher covalently attached to the 5' and 3' ends of an oligo typically 20 to 30 bases in length. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridization to its complementary sequence. Dual-Labeled BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences. BHQ probes are offer delivery of the probe and primers in separate individual tubes. We also offer ValuMix for gene expression and qPCR which delivers one probe and two primers in a single tube. ValumixPrimer quantities can be flexibly adjusted around a specified probe amount of either 0.5 nmol, 5 nmol, or 20 nmol delivered. Probe to primer ratios can range anywhere from 1:1 to 1:4.5 according to your preference. ValuMix is an economical option providing 0.5 nmol of FAM-BHQ probe mixed with primers for only $79. See the product listing tab for full range of options. Design Concerns for BHQ probes Please be aware that sequences longer than 30 bases are unlikely to have efficient quenching as an end-labeled probe. For such situations please consider positioning the quencher internally instead. Conversely, sequences shorter than 20 bases may have been designed as a Minor Groove Binder (MGB™) probe and are unlikely to bind at the proper temperature of 70 °C. Please consider ordering such a sequence as a BHQplus® probe instead, or else contact info@genecompany.com with any questions regarding probe design. NOTE: Building Probes with an Internal Quencher (Currently available with FAM only) Adding an internal modification annotated by T(...) indicates that a Thymidine base is attached to the internal modification. Please be aware that an additional T base will be introduced into the sequence when adding such an internal modification. Building Probes with Wobble/Degenerate Bases (What are Wobbles?) To ensure the most efficient processing of your oligo, use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc. Best DEAL: ValuProbe™ BHQ Probe - 10 nanomoles of FAM-BHQ Probe typically ships in 2 business days Instrument CALIBRATION: FAM, CAL Fluor and Quasar calibration dyes are available here for online purchase Learn MORE: Applying dual-labeled probes in Multiplexing qPCR, Gene Expression, and SNP Genotyping assays The difference between FRET and static quenching Assay DESIGN: Try RealTimeDesign™ Software, a FREE, qPCR assay design program. Design your Dual-Labeled BHQ probes and primers online today!

https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhq-probes#?tab=product-info 双标记的BHQ探针是一种传统的线性FRET探针，其将荧光基团和淬灭基团共价连接到通常长度为20至30个碱基的寡核苷酸的5'和3'末端。荧光通过Taq聚合酶的5'外切核酸酶活性释放，Taq聚合酶在探针与其互补序列杂交时切割荧光染料。双标记BHQ探针是检测特异性靶序列的存在和定量的理想选择。 BHQ探针提供两种形式： 1）探针和引物在分开在单独管中形式； 2）用于基因表达和qPCR的ValuMix形式，即在单管中包含一个探针和两个引物的混合物。 ValumixPrimer的数量可以根据指定的探针量进行灵活地调整，如0.5 nmol，5 nmol或20 nmol。探针与引物的比率可以根据您的喜好在1：1至1：4.5的范围内。ValuMix是一个经济的选择，LGC Biosearch以经济实惠的价格提供0.5 nmol超小包装FAM-BHQ探针引物混合物。有关完整的选择范围，请参阅官网产品列表选项卡。 BHQ探针设计注意事项： 双标记探针作为一种末端标记探针，当序列长于30个碱基时会影响荧光淬灭效率。对于这种情况，请考虑BHQnova双猝灭探针（https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhqnova-probes）或将猝灭剂放在探针中间位置设计具有内部猝灭的探针（该形式当前仅提供FAM标记）。相反，短于20个碱基的序列可以被设计为小沟结合剂（MGB）探针，并且不可能在70℃的适当温度下结合。请考虑订购BHQplus®探针，或者联系基因有限公司技术人员咨询有关探针设计的任何问题。 注意： 1. 设计具有内部猝灭的探针（当前仅提供FAM） 2. 添加由T（...）注释的内部修饰表示胸腺嘧啶碱基连接到内部修饰。请注意，在添加此类内部修饰时，将在序列中引入另外的T碱基。 3. 具有不确定/降解碱基的序列如何设计探针，请参考https://www.biosearchtech.com/support/faqs/custom-oligonucleotides-modifications/what-are-wobbles 4. 为了确保寡核苷酸能被有效识别，当在序列输入框中输入不确定/降解碱基时请使用IUPAC代码，而非括号，如：（A / G），（A / G / C / T）等。 Dual-Labeled BHQ Probes are traditional, linear, FRET probes incorporating a fluorophore and quencher covalently attached to the 5' and 3' ends of an oligo typically 20 to 30 bases in length. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridization to its complementary sequence. Dual-Labeled BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences. BHQ probes are offer delivery of the probe and primers in separate individual tubes. We also offer ValuMix for gene expression and qPCR which delivers one probe and two primers in a single tube. ValumixPrimer quantities can be flexibly adjusted around a specified probe amount of either 0.5 nmol, 5 nmol, or 20 nmol delivered. Probe to primer ratios can range anywhere from 1:1 to 1:4.5 according to your preference. ValuMix is an economical option providing 0.5 nmol of FAM-BHQ probe mixed with primers for only $79. See the product listing tab for full range of options. Design Concerns for BHQ probes Please be aware that sequences longer than 30 bases are unlikely to have efficient quenching as an end-labeled probe. For such situations please consider positioning the quencher internally instead. Conversely, sequences shorter than 20 bases may have been designed as a Minor Groove Binder (MGB™) probe and are unlikely to bind at the proper temperature of 70 °C. Please consider ordering such a sequence as a BHQplus® probe instead, or else contact info@genecompany.com with any questions regarding probe design. NOTE: Building Probes with an Internal Quencher (Currently available with FAM only) Adding an internal modification annotated by T(...) indicates that a Thymidine base is attached to the internal modification. Please be aware that an additional T base will be introduced into the sequence when adding such an internal modification. Building Probes with Wobble/Degenerate Bases (What are Wobbles?) To ensure the most efficient processing of your oligo, use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc. Best DEAL: ValuProbe™ BHQ Probe - 10 nanomoles of FAM-BHQ Probe typically ships in 2 business days Instrument CALIBRATION: FAM, CAL Fluor and Quasar calibration dyes are available here for online purchase Learn MORE: Applying dual-labeled probes in Multiplexing qPCR, Gene Expression, and SNP Genotyping assays The difference between FRET and static quenching Assay DESIGN: Try RealTimeDesign™ Software, a FREE, qPCR assay design program. Design your Dual-Labeled BHQ probes and primers online today!

https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhq-probes#?tab=product-info 双标记的BHQ探针是一种传统的线性FRET探针，其将荧光基团和淬灭基团共价连接到通常长度为20至30个碱基的寡核苷酸的5'和3'末端。荧光通过Taq聚合酶的5'外切核酸酶活性释放，Taq聚合酶在探针与其互补序列杂交时切割荧光染料。双标记BHQ探针是检测特异性靶序列的存在和定量的理想选择。 BHQ探针提供两种形式： 1）探针和引物在分开在单独管中形式； 2）用于基因表达和qPCR的ValuMix形式，即在单管中包含一个探针和两个引物的混合物。 ValumixPrimer的数量可以根据指定的探针量进行灵活地调整，如0.5 nmol，5 nmol或20 nmol。探针与引物的比率可以根据您的喜好在1：1至1：4.5的范围内。ValuMix是一个经济的选择，LGC Biosearch以经济实惠的价格提供0.5 nmol超小包装FAM-BHQ探针引物混合物。有关完整的选择范围，请参阅官网产品列表选项卡。 BHQ探针设计注意事项： 双标记探针作为一种末端标记探针，当序列长于30个碱基时会影响荧光淬灭效率。对于这种情况，请考虑BHQnova双猝灭探针（https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhqnova-probes）或将猝灭剂放在探针中间位置设计具有内部猝灭的探针（该形式当前仅提供FAM标记）。相反，短于20个碱基的序列可以被设计为小沟结合剂（MGB）探针，并且不可能在70℃的适当温度下结合。请考虑订购BHQplus®探针，或者联系基因有限公司技术人员咨询有关探针设计的任何问题。 注意： 1. 设计具有内部猝灭的探针（当前仅提供FAM） 2. 添加由T（...）注释的内部修饰表示胸腺嘧啶碱基连接到内部修饰。请注意，在添加此类内部修饰时，将在序列中引入另外的T碱基。 3. 具有不确定/降解碱基的序列如何设计探针，请参考https://www.biosearchtech.com/support/faqs/custom-oligonucleotides-modifications/what-are-wobbles 4. 为了确保寡核苷酸能被有效识别，当在序列输入框中输入不确定/降解碱基时请使用IUPAC代码，而非括号，如：（A / G），（A / G / C / T）等。 Dual-Labeled BHQ Probes are traditional, linear, FRET probes incorporating a fluorophore and quencher covalently attached to the 5' and 3' ends of an oligo typically 20 to 30 bases in length. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridization to its complementary sequence. Dual-Labeled BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences. BHQ probes are offer delivery of the probe and primers in separate individual tubes. We also offer ValuMix for gene expression and qPCR which delivers one probe and two primers in a single tube. ValumixPrimer quantities can be flexibly adjusted around a specified probe amount of either 0.5 nmol, 5 nmol, or 20 nmol delivered. Probe to primer ratios can range anywhere from 1:1 to 1:4.5 according to your preference. ValuMix is an economical option providing 0.5 nmol of FAM-BHQ probe mixed with primers for only $79. See the product listing tab for full range of options. Design Concerns for BHQ probes Please be aware that sequences longer than 30 bases are unlikely to have efficient quenching as an end-labeled probe. For such situations please consider positioning the quencher internally instead. Conversely, sequences shorter than 20 bases may have been designed as a Minor Groove Binder (MGB™) probe and are unlikely to bind at the proper temperature of 70 °C. Please consider ordering such a sequence as a BHQplus® probe instead, or else contact info@genecompany.com with any questions regarding probe design. NOTE: Building Probes with an Internal Quencher (Currently available with FAM only) Adding an internal modification annotated by T(...) indicates that a Thymidine base is attached to the internal modification. Please be aware that an additional T base will be introduced into the sequence when adding such an internal modification. Building Probes with Wobble/Degenerate Bases (What are Wobbles?) To ensure the most efficient processing of your oligo, use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc. Best DEAL: ValuProbe™ BHQ Probe - 10 nanomoles of FAM-BHQ Probe typically ships in 2 business days Instrument CALIBRATION: FAM, CAL Fluor and Quasar calibration dyes are available here for online purchase Learn MORE: Applying dual-labeled probes in Multiplexing qPCR, Gene Expression, and SNP Genotyping assays The difference between FRET and static quenching Assay DESIGN: Try RealTimeDesign™ Software, a FREE, qPCR assay design program. Design your Dual-Labeled BHQ probes and primers online today!

https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhq-probes#?tab=product-info 双标记的BHQ探针是一种传统的线性FRET探针，其将荧光基团和淬灭基团共价连接到通常长度为20至30个碱基的寡核苷酸的5'和3'末端。荧光通过Taq聚合酶的5'外切核酸酶活性释放，Taq聚合酶在探针与其互补序列杂交时切割荧光染料。双标记BHQ探针是检测特异性靶序列的存在和定量的理想选择。 BHQ探针提供两种形式： 1）探针和引物在分开在单独管中形式； 2）用于基因表达和qPCR的ValuMix形式，即在单管中包含一个探针和两个引物的混合物。 ValumixPrimer的数量可以根据指定的探针量进行灵活地调整，如0.5 nmol，5 nmol或20 nmol。探针与引物的比率可以根据您的喜好在1：1至1：4.5的范围内。ValuMix是一个经济的选择，LGC Biosearch以经济实惠的价格提供0.5 nmol超小包装FAM-BHQ探针引物混合物。有关完整的选择范围，请参阅官网产品列表选项卡。 BHQ探针设计注意事项： 双标记探针作为一种末端标记探针，当序列长于30个碱基时会影响荧光淬灭效率。对于这种情况，请考虑BHQnova双猝灭探针（https://www.biosearchtech.com/products/qpcr-and-snp-genotyping/bhqnova-probes）或将猝灭剂放在探针中间位置设计具有内部猝灭的探针（该形式当前仅提供FAM标记）。相反，短于20个碱基的序列可以被设计为小沟结合剂（MGB）探针，并且不可能在70℃的适当温度下结合。请考虑订购BHQplus®探针，或者联系基因有限公司技术人员咨询有关探针设计的任何问题。 注意： 1. 设计具有内部猝灭的探针（当前仅提供FAM） 2. 添加由T（...）注释的内部修饰表示胸腺嘧啶碱基连接到内部修饰。请注意，在添加此类内部修饰时，将在序列中引入另外的T碱基。 3. 具有不确定/降解碱基的序列如何设计探针，请参考https://www.biosearchtech.com/support/faqs/custom-oligonucleotides-modifications/what-are-wobbles 4. 为了确保寡核苷酸能被有效识别，当在序列输入框中输入不确定/降解碱基时请使用IUPAC代码，而非括号，如：（A / G），（A / G / C / T）等。 Dual-Labeled BHQ Probes are traditional, linear, FRET probes incorporating a fluorophore and quencher covalently attached to the 5' and 3' ends of an oligo typically 20 to 30 bases in length. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridization to its complementary sequence. Dual-Labeled BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences. BHQ probes are offer delivery of the probe and primers in separate individual tubes. We also offer ValuMix for gene expression and qPCR which delivers one probe and two primers in a single tube. ValumixPrimer quantities can be flexibly adjusted around a specified probe amount of either 0.5 nmol, 5 nmol, or 20 nmol delivered. Probe to primer ratios can range anywhere from 1:1 to 1:4.5 according to your preference. ValuMix is an economical option providing 0.5 nmol of FAM-BHQ probe mixed with primers for only $79. See the product listing tab for full range of options. Design Concerns for BHQ probes Please be aware that sequences longer than 30 bases are unlikely to have efficient quenching as an end-labeled probe. For such situations please consider positioning the quencher internally instead. Conversely, sequences shorter than 20 bases may have been designed as a Minor Groove Binder (MGB™) probe and are unlikely to bind at the proper temperature of 70 °C. Please consider ordering such a sequence as a BHQplus® probe instead, or else contact info@genecompany.com with any questions regarding probe design. NOTE: Building Probes with an Internal Quencher (Currently available with FAM only) Adding an internal modification annotated by T(...) indicates that a Thymidine base is attached to the internal modification. Please be aware that an additional T base will be introduced into the sequence when adding such an internal modification. Building Probes with Wobble/Degenerate Bases (What are Wobbles?) To ensure the most efficient processing of your oligo, use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc. Best DEAL: ValuProbe™ BHQ Probe - 10 nanomoles of FAM-BHQ Probe typically ships in 2 business days Instrument CALIBRATION: FAM, CAL Fluor and Quasar calibration dyes are available here for online purchase Learn MORE: Applying dual-labeled probes in Multiplexing qPCR, Gene Expression, and SNP Genotyping assays The difference between FRET and static quenching Assay DESIGN: Try RealTimeDesign™ Software, a FREE, qPCR assay design program. Design your Dual-Labeled BHQ probes and primers online today!