KAPA KK4824-Library Quantification Kit 文库定量试剂

KAPA KK4824-Library Quantification Kit 文库定量试剂

适用Illumina文库

KAPA Biosystems是一家美国生物公司,成立于2006年,其生产基地位于南非开普敦。KAPA Biosystems致力于研发、生产下一代PCR(Next-Generation PCR)所需的生物酶。目前其分子进化平台开发的产品已被广泛应用于DNA扩增、二代测序、分子诊断等领域。

KAPA KK4824-Library Quantification Kit 文库定量试剂

英文名称:KAPA Library Quantification Kits for Illumina platforms

中文名称:illumina测序文库定量试剂盒

KAPA 文库定量试剂盒(illumina平台)特别设计用于Illumina测序文库的准确和可重复的定量,包含qPCR定量所需所有试剂。试剂盒含有新型KAPA SYBR FAST DNA聚合酶,保证效率的前提下,能定量平均片段长度高达1kb的Illumina文库。

产品特点

试剂盒适用序列GC含量范围宽泛,平均片段长度可达1 kb。

除了NGS文库定量之外,该试剂盒还可用于检测Illumina文库制备过程中的污染情况。

文库定量分析包括重复的移液步骤,容易自动化。

经过严格测试,以确保zui小的批次间差异。

产品应用和兼容性

KAPA Library定量试剂盒(illumina平台)特别设计用于Illumina测序文库的准确和可重复的定量。适用文库不限类型、构建方式和illumina测序仪型号,只要浓度> 0.0002 pM,并且含有与试剂盒中引物互补的序列(Illumina P5 and P7流动池寡核苷酸序列),都可以使用该试剂盒进行定量。

KAPA Library Quantification Kits for Illumina platforms are suitable for the quantification of any NGS library (prepared for Illumina sequencing) which contains the P5 and P7 flow cell sequence motifs. Libraries constructed using full-length universal or indexed TruSeq™ adapters can be quantified after adapter ligation. Libraries prepared with partial or stem-loop adapters can only be quantified with this kit after library amplification, during which indexes (and flow cell sequences) are added. To ensure compatibility, compare the adapter sequences used during library preparation with the library quantification primer sequences in Product Description.

Ensure that the correct KAPA SYBR FAST qPCR Master Mix is used, in accordance with the reference dye requirements (if any) of the qPCR instrument to be used for library quantification.

产品组份

 

 

 

 

 

 

 

 

适用于Illumina平台的KAPA Library定量试剂盒包含所有含P5和P7流动池序列的Illumina文库进行qPCR定量所需的完整试剂。

完全试剂盒包含标准DNA,引物混合物,qPCR混合液

试剂盒包含:

•文库定量DNA标准品 1 – 6 (a 10-fold dilution series of a linear, 452 bp template)

•文库定量引物Primer Premix (10X), 引物序列:

Primer 1: 5′-AAT GAT ACG GCG ACC ACC GA-3′

Primer 2: 5′-CAA GCA GAA GAC GGC ATA CGA-3′

•KAPA SYBR® FAST qPCR Master Mix (2X),各种passive reference染料。

该试剂盒含有新型KAPA SYBR FAST DNA聚合酶,通过定向进化过程设计用于高性能SYBR Green I型qPCR。改造的聚合酶以相似的效率扩增不同DNA片段的能力使得能够使用通用标准来可靠地量化平均片段长度高达1kb的所有Illumina文库,而不管文库类型或GC含量如何。

KAPA SYBR FAST是一种抗体介导的热启动DNA聚合酶制剂。

使用方法

1. 使用KAPA SYBR FAST qPCR Master Mix和引物(靶向Illumina P5和P7流动池序列)

通过qPCR法扩增DNA标准品(6个,预稀释)和稀释的文库样品来进行文库定量。

2. 绘制每个DNA标准品的平均Cq值与log10(浓度:pM)的关系曲线,以产生标准曲线。使用定量对照标准曲线计算稀释的文库样品的浓度。

注意:高通量NGS管路强烈建议使用自动化液体处理系统。有关更多信息,请参阅KAPA库量化技术指南。

Library quantification is performed by amplifying the set of six pre-diluted DNA Standards and diluted library samples by qPCR, using the KAPA SYBR FAST qPCR Master Mix and primers targeting the Illumina P5 and P7 flow cell oligo sequences. The average Cq score for each DNA Standard is plotted against log10(concentration in pM) to generate a standard curve. The concentrations of diluted library samples are then calculated against the standard curve, using absolute quantification.