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GeneRead Library Quant System

用于qPCR定量检测NGS文库
  • 以芯片规格提供预分装的即用型、梯度稀释DNA标准品
  • 自动计算文库浓度
  • 实验方案简便、灵敏度高、动态范围宽
  • 与Illumina和Ion Torrent/Proton NGS平台兼容
  • 与多数qPCR仪兼容
GeneRead Library Quant System为NGS文库制备提供简便的解决方案,确保NGS实验获得可靠结果。GeneRead Library Quant System采用real-time PCR技术,特异性定量检测两端有测序接头的DNA分子。
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Available Products
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qPCR arrays (96-well format) for quantifying Illumina libraries
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333304 QSIL-003 varies
qPCR arrays (384-well format) for quantifying Illumina libraries
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333304 QSIL-3003 varies
qPCR arrays (96-well format) for quantifying Ion Torrent libraries
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333304 QSIT-003 varies
qPCR arrays (384-well format) for quantifying Ion Torrent libraries
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333304 QSIT-3003 varies
qPCR assays for quantifying Illumina libraries
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qPCR assays for quantifying Ion Torrent libraries
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333314 QSTF-ITZ varies
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GeneRead Library Quant System适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


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GeneRead Library Quant Array工作流程
将各样本DNA文库与GeneRead qPCR SYBR® Green Mastermix混合,一式三份加入GeneRead Library Quant Array,之后用qPCR进行分析。将原始CT值复制到数据分析Excel模板中,软件会自动计算文库浓度或稀释因子。
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GeneRead Library Quant System能够定量检测低浓度文库,甚至是浓度低于传统分光光度法的检测阈限。
GeneRead Library Quant Array灵敏性高,动态范围宽泛,能够定量检测NGS-L1和NGS-L2文库[A]。相反,Agilent's High Sensitivity DNA Kit(与Agilent 2100 BioAnalyzer配套使用)只能定量检测NGS-L1文库;对于这个试剂盒,NGS-L2浓度太低,无法进行定量检测[B]。
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GeneRead Library Quant System的原理。
DNA标准品的梯度稀释液(1个10倍梯度稀释液)能够获得标准曲线,样本文库浓度落在标准曲线上。
Performance
使用两种浓度的NGS文库,将GeneRead Library Quant System的灵敏度和动态范围与Agilent's High Sensitivity DNA Kit进行对比。第一个文库使用这两种试剂盒皆可进行定量检测,但浓度较低的第二个文库低于Agilent's 2100 BioAnalyzer的最低检测阈限。相反,GeneRead Library Quant Array能够对这两种不同浓度的文库进行成功的定量检测(参见GeneRead Library Quant System enables quantification of libraries with concentrations below the detection limit of conventional methods)。
Principle

准确定量文库对于NGS平台的高效使用十分重要。但是,当前的分光广度法只能检测总核酸浓度。GeneRead Library Quant System则采用real-time PCR技术,仅定量检测两端有接头的DNA分子。只有两端有接头的DNA分子才能在乳液PCR(Ion Torrent)或桥式PCR(Illumina)中得到扩增,因此此方法能够准确定量检测可扩增的文库DNA。real-time PCR灵敏性高,能够定量检测低浓度文库,甚至是浓度低于传统分光光度法的检测阈限。

GeneRead Library Quant System提供管装和芯片规格,经过优化,适合配合GeneRead qPCR SYBR Green Mastermixes使用,具有出色的灵敏性和宽泛的动态范围。两种规格都提供适合Illumina或Ion Torrent平台的DNA标准品。芯片规格提供分装的5个浓度的10倍梯度稀释标准品,混合有PCR引物,一式三份。确保您的样本文库在检测范围之中(参见Principle of GeneRead Library Quant System)。

Procedure

芯片工作流程(参见GeneRead Library Quant Array workflow)使用两个10倍稀释的样本文库,确保其浓度在梯度稀释的DNA标准品检测范围内。将样本与Generead qPCR SYBR Green Mastermix混合,等分为三份,加入芯片中。进行PCR,将原始CT数据导出至提供的数据分析Excel文件中,进行文库浓度的自动计算(Illumina)或模板稀释因子计算(Ion Torrent/Proton)。

管装标准品的工作流程与芯片的工作流程相似,仅有少数不同。先制备5个10倍梯度稀释的DNA标准品,以及2个10倍梯度稀释的样本文库。这样可确保文库的浓度在标准品稀释液范围之内。下一步,将稀释的DNA标准品和样本文库与提供能够的PCR试剂和相应的GeneRead qPCR SYBR Green Mastermix混合。进行PCR,将原始CT数据导出至提供的数据分析Excel文件中,进行文库浓度的自动计算(Illumina)或模板稀释因子计算(Ion Torrent/Proton)。

Applications
GeneRead Library Quant Array和GeneRead Library Quant Kit能够在使用Illumina或Ion Torrent/Proton NGS平台测序前,快速、准确的检测样本文库浓度。

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分析软件 (8)
New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) or GeneRead Library Quant Array for 384-well plates for Illumina libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) or GeneRead Library Quant Array for 384-well plates for Ion Torrent libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) or GeneRead Library Quant Array for 96-well plates for Illumina libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) or GeneRead Library Quant Array for 96-well plates for Ion Torrent libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) or GeneRead Library Quant Array for Rotor-Disc 100 rings for Illumina libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) or GeneRead Library Quant Array for Rotor-Disc 100 rings for Ion Torrent libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) for 72-well Rotor rings for Illumina libraries
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New version – Excel-based data analysis template for use with GeneRead Library Quant Kit (assay format) for 72-well Rotor rings for Ion Torrent libraries
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产品介绍与指南 (2)
Accelerate your NGS performance through Sample to Insight solutions
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Accelerate your NGS performance through Sample to Insight solutions
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