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PyroMark Q24 Advanced Reagents

适用于在长序列测序中利用焦磷酸测序实现先进的甲基化和突变定量
  • 先进的技术、软件和化学试剂,可用于长序列分析
  • 连续CpG或CpN位点的定量甲基化分析
  • 更优化的任意序列位置序列差异定量
PyroMark Q24 Advanced Reagents采用先进的焦磷酸测序试剂,可提供比以往更佳的实时测序检测和定量。PyroMark Q24 Advanced Reagents专为PyroMark Q24 Advanced优化,适用于分析各种类型的序列差异,尤其是CpG或CpN位点的DNA甲基化、复杂突变或用于从头测序应用(如微生物分型)。长序列测序(如从头测序或甲基化分析)可能需要更多量的核苷酸,我们推荐使用PyroMark Q24 Advanced CpG Reagents。
Cat No./ID: 970902
PyroMark Q24 Advanced Reagents (4 x 24)
Enzyme mix, substrate mix, nucleotides, and buffers for Pyrosequencing assays; intended for use with PyroMark Q24 Advanced
Cat No./ID: 970922
PyroMark Q24 Advanced CpG Reagents (4 x 24)
Enzyme mix, substrate mix, nucleotides, and buffers for CpG and long read Pyrosequencing assays; intended for use with PyroMark Q24 Advanced

PyroMark Q24 Advanced Reagents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。


Performance
更长的焦磷酸测序运行和更高的准确度
PyroMark Q24 Advanced采用了改进的化学试剂和仪器操作算法,大大增加了读取长度和碱基检出功能的准确度,可轻松实现较长的从头测序运行分析。此前,读取长度受到测序反应中的背景峰和较低的光信号的限制。已升级的PyroMark Q24 Advanced化学试剂和算法可降低该背景峰,从而提高读取长度和可靠性。根据待分析序列的不同,只需一次PyroMark Q24 Advanced反应即可获得高度准确的140个或更多碱基的读取长度 (参见Long de novo sequencing runs)。

任意位置的更先进的甲基化分析
PyroMark Q24 Advanced可在任意序列位置实现改进的长片段序列甲基化定量。以前,远离测序引物的甲基化位点分析是无法确定的,但现在利用更长的读取长度和更高的准确度,可在整个测序运行中实现高度可靠的甲基化定量(参见Analysis of 16 CpG sites in a long sequence run)。

改进的同聚物测序准确度
DNA甲基化分析中的亚硫酸氢盐转化可导致核苷酸序列中频繁的poly T延伸,以前,由于这些位点的光信号定量的不确定性,在这种T同聚物之后直接进行CpG位点分析面临诸多难题。PyroMark Q24 Advanced具有更高的准确度,可在8个T核苷酸链之后甚至之中实现可靠的CpG甲基化定量(参见Improved methylation quantification in homopolymers)。

长片段序列的突变分析
PyroMark Q24 Advanced还可利用单个Assay实现可靠的多态性定量。由于单核苷酸多态性(SNP)及其他突变通常不相邻,因此传统的焦磷酸测序化学方法一般需要分析针对每个突变位点的单独的Assay。全新的PyroMark Q24 Advanced化学方法可实现更长片段分析,在一次运行中实现多个突变或SNP的可靠分析(参见Quantitative mutation analysis in long sequence runs)。 
Principle
焦磷酸测序技术采用了合成测序原理,可在数分钟内提供序列定量数据。PyroMark Q24 Advanced Reagents和PyroMark Q24 Advanced CpG Reagents采用升级的焦磷酸测序试剂,可分析复杂突变、表观遗传学甲基化研究、抗性分型和微生物鉴定分析。这些试剂专为配套PyroMark Q24 Advanced仪器经过优化。
Procedure
快速方便的样本制备
在焦磷酸测序之前,先生成生物素化PCR产物。该生物素化PCR产物可与链霉亲和素包被的Sepharose微珠结合,利用Vacuum Workstation上的Vacuum Tool可捕获微珠,彻底冲洗并变性,生成适用于焦磷酸测序的单链DNA。将上述模板DNA加至包含测序引物的焦磷酸反应板中,引物退火后,将反应板置于PyroMark仪器上。PyroMark Q24 Advanced Reagents或PyroMark Q24 Advanced CpG Reagents包含焦磷酸测序反应所需的酶、核苷酸和底物;参照软件提供的体积,将试剂加至分装装置内,同样置于仪器内供焦磷酸测序使用。
Applications
焦磷酸测序在各学科的研究应用中发挥了愈来愈重要的作用。不论是检查病原体中的抗药性的产生、基因表达调控中表观遗传学DNA甲基化的作用、家畜中特定表型的遗传标志物,还是线粒体DNA法医样本中的多态性,PyroMark Q24 Advanced均可提供功能强大且通用的基因和表观遗传学变异分析。此外,由于焦磷酸测序整合了序列检测和定量,提高的分析分辨率可促进新发现的诞生。

产品对比
PyroMark Q24 Advanced PyroMark Q24
通量 1–24 个样本 1–24个样本
体积 25 µl 25 µl
PCR产物需要量 5–10 µl
(~0.5–3 pmol产物)
5–10 µl
(~0.5–3 pmol产物)
读长(取决于Assay和序列) 10–140 bp以上 10–80 bp
应用软件 PyroMark Q24 Advanced SW
(需要固件1.5.6903或更高)
PyroMark Q24 SW 2.0
软件功能 SEQ(从头测序)
CpG/CpN甲基化
SNP
AQ
SQA (从头测序)
CpG甲基化
AQ/SNP
主要应用 复杂突变分析
表观遗传学(CpG和CpN分析)
耐药性分析和微生物鉴定
突变分析
耐药性分析
兼容试剂 PyroMark Q24 Advanced Reagents
PyroMark Q24 Advanced CpG Reagents
PyroMark Q24 Gold Reagents
灵敏度 2%的突变
98% wt
2%的突变
98% wt

重要信息:PyroMark Q24 Advanced Reagents和PyroMark Q24 Advanced CpG Reagents与PyroMark Q24 Software不可配套使用。PyroMark Gold Q24 Reagents不可与PyroMark Q24 Advanced Software配套使用。试剂和软件的错误搭配会导致试剂分配错误,检测结果出错。

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下载文件 (11)
Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0003
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0004
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0005
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0006
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0007
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0008
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0009
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0010
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0011
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0012
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Instrument methods file for use with PyroMark Q24 Advanced Software, PyroMark Q24 Advanced Reagents or PyroMark Q24 Advanced CpG Reagents, and PyroMark Q24 Cartridges marked with method 0013
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应用说明 (1)
A Pyrosequencing method for the forensic identification of tissue source using QIAGEN’s PyroMark Q24 Advanced system.
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