NGSBIO Illumina 平台 DNA 标准品

​特点​

1. 可见的蓝色预混液，有助于提高加样精确度和准确性

2. 使用qPCR在测序前快速、准确地定量文库

3. 对多种样品类型、浓度、片段大小和GC含量的文库均能提供一致的定量结果

4. 对所有文库使用单一延伸时间

5. 适用于手动和自动化工作流程

6. 特异性定量仅可用于NGS测序的DNA分子

7. 采用抗体介导的热启动技术，确保所有反应同步开始

8. 兼容所有Illumina测序仪和qPCR平台

9. 提供可见的蓝色qPCR预混液版本

10. 使用在线NGSBIO文库定量工具轻松计算文库浓度

概述

NGSBIO 文库定量试剂盒包含了对为Illumina NGS系统制备的文库进行精准、灵敏定量所需的所有组分。该试剂盒采用qPCR方法特异性定量连接了接头的DNA分子，从而确保获得最佳簇密度，以改进测序效率和数据质量。​

NGSBIO 文库定量试剂盒提供了一种可靠的基于qPCR的方法，用于定量为Illumina NGS系统制备的文库。该试剂盒包含5个DNA标准品、特异性针对Illumina P5和P7接头序列的引物以及qPCRBIO SyGreen® Mix预混液。先进的qPCR缓冲液系统采用高通量智能筛选技术开发，确保能高效扩增所有文库，包括那些GC或AT含量偏高的文库。试剂盒还提供便捷的文库稀释缓冲液。该试剂盒也可与VeriFi™ 文库扩增预混液配合使用。

The NGSBIO Library Quant Kit offers a reliable qPCR-based method for the quantification of libraries prepared for Illumina NGS systems. The kit includes 5 DNA standards, primers specific to the P5 and P7 Illumina adapter sequences and qPCRBIO SyGreen® Mix. The advanced qPCR buffer system has been developed using our high-throughput smart screen technology to ensure efficient amplification of all your libraries, including those that are GC or AT-rich. The kit is also supplied with a convenient library dilution buffer

​精准的文库定量​

qPCR被认为是定量NGS文库的最佳方法，因为它只测量那些可作为文库扩增和簇生成模板的接头连接分子。NGSBIO 文库定量试剂盒能够实现高度精准的定量，这对于获得最佳簇密度和更高测序效率至关重要。

文库定量可在接头连接后的任何时间进行，并且应在簇生成之前使用。为方便准确地进行数据分析，我们推荐使用NGSBIO文库定量工具。高估文库浓度可能导致簇密度不足，而低估文库浓度则可能导致簇密度过高达到饱和。要了解有关最佳簇密度的更多信息，请参阅您Illumina测序仪的操作说明。

qPCR is considered the best method for quantifying NGS libraries as it only measures adapter-bound molecules that can be used as templates for library amplification and cluster generation. The NGSBIO Library Quant Kit enables highly accurate quantification crucial for optimal cluster densities and greater sequencing efficiency. 

Library quantification can be used at any time after adapter ligation and should always be used prior to cluster generation. For easy and accurate data analysis we recommend using the . Overestimating library concentration can result in insufficient cluster density and underestimating library concentration can result in a high, saturating cluster density. To learn more about optimal cluster density, consult the instructions of your Illumina machine.

宽动态范围​

试剂盒提供的DNA标准品经过精确定量，即开即用，覆盖5个数量级（从2 pM 到 0.2 fM）。该试剂盒适用于甚至低浓度文库的定量，包括无需PCR扩增步骤构建的文库。

​通用型试剂盒​

NGSBIO 文库定量试剂盒兼容所有qPCR平台，并经过优化，可在多种样品类型、片段大小（高达1000bp）、浓度和GC含量范围内提供一致且可重复的文库定量结果。使用我们的qPCR选择工具查找适合您仪器的ROX变体。

为方便准确地进行数据分析，我们推荐使用NGSBIO文库定量工具。

​使用NGSBIO Illumina平台文库定量试剂盒与竞争对手试剂盒（NEB, KAPA）及Qubit荧光计获得的NGS文库定量值对比分析图​

• ​高一致性和重复性：​​ 使用NGSBIO文库定量试剂盒（紫色）、NEBNext®文库定量试剂盒（橙色）和KAPA文库定量试剂盒（绿色）对16个接头连接文库进行定量。NGSBIO文库定量试剂盒显示重复样品的变异更小，一致性更高。定量结果与两个领先的NGS文库定量试剂盒制造商的产品结果相当。每个图表右下角的数字代表使用Invitrogen的Qubit荧光计测得的dsDNA浓度。

• ​High consistency and reproducibility of quantification: 16 adapter-ligated libraries are quantified using NGSBIO Library Quant Kit (purple), NEBNext® Library Quant Kit (orange) and KAPA Library Quantification Kit (green). The NGSBIO Library Quant Kit shows less spread and greater consistency among replicates. The quantification results are within those obtained by two leading manufacturers of NGS library quantification kits. The number on the bottom right corner of each graph represents the concentration of dsDNA obtained using a Qubit Fluorometer from Invitrogen.

使用NGSBIO Illumina平台文库定量试剂盒生成的扩增曲线和标准曲线图​

• ​扩增曲线和标准曲线：​​ 将一个接头连接文库样品（紫色）与NGSBIO文库定量试剂盒提供的六个标准品模板（灰色）同时运行（图a）。将扩增曲线的Ct值对标准品模板浓度的对数作图。通过标准品点拟合一条线性曲线（图b）。然后根据未知样品在曲线上的位置计算其浓度。

• ​Amplification curves and standard curve: An adapter-ligated library sample (purple) is run alongside six standard templates (grey) provided in the NGSBIO Library Quant Kit (a). The Cts of the amplification curves are plotted against the log of the concentration of the standard templates. A linear curve is fitted through the standards (b). The concentration of the unknown sample is then calculated from its position on the curve.