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miScript SYBR® Green PCR Kit

配合miScript PCR System,使用real-time PCR方法检测miRNA

Features

  • 灵敏并特异性定量检测miRNA
  • 利用单一cDNA定量检测多种miRNA
  • 利用同一cDNA定量检测miRNA和mRNA
  • 包含ROX染料,可用于标准化荧光结果

Product Details

miScript SYBR Green PCR Kit是miScript PCR System不可缺少的部分,用于定量检测miRNA。该试剂盒基于经验证的QuantiTect SYBR Green Kits技术。miScript II RT Kit得到的cDNA作为模板,配合miScript SYBR Green PCR Kit和PCR芯片(miRNome或Pathway-Focused miScript miRNA PCR Arrays)或适当的检测(用于miRNA或其它非编码RNA的miScript Primer Assays、用于miRNA前体的miScript Precursor Assays或用于mRNA的QuantiTect Primer Assays)进行real-time PCR。

Performance

miScript SYBR Green PCR Kit配合miScript PCR System的组件,灵敏和特异性的检测和定量miRNA。miScript PCR System灵敏度高、动态范围广。持续高的扩增效率,确保使用∆∆CT方法进行多个miRNAs表达分析结果的准确性(参见" miRNA expression in colorectal cancer")。其卓越的性能使得初次尝试时即可获得高度重复性结果。
See figures

Principle

miScript PCR System配合SYBR Green-based real-time PCR,灵敏和特异性的定量和分析miRNA。miScript PCR System可以完成从RNA中合成cDNA和后续的real-time PCR检测miRNAs涉及的所有步骤。

miScript PCR System包含以下组分:

miScript II RT Kit,可简单、单步合成cDNA。单个cDNA合成反应可用于检测数以百计的miRNAs。双缓冲体系满足使用real-time PCR进行miRNA定量的独特要求。
miScript SYBR Green PCR Kit,包含QuantiTect SYBR Green PCR Master Mix和反向引物miScript Universal Primer,配合miScript Primer Assay或miScript miRNA PCR Array进行miRNAs检测。
miScript Primer Assay或miScript Precursor Assay,包含QuantiTect SYBR Green PCR Master Mix和反向引物miScript Universal Primer,配合miScript Primer Assay或miScript miRNA PCR Array进行miRNAs检测。
miScript miRNA PCR Array,miRNome或Pathway-Focused miScript miRNA PCR Arrays被预先设定,单次使用的PCR芯片可快速分析成熟miRNAs。
miScript miRNA PCR Array数据分析仪,免费的基于网络的工具,简化了用于miScript miRNA PCR Arrays相对定量的ΔΔCT方法。
Real-time PCR检测miRNA

miScript SYBR Green PCR Kit包含QuantiTect SYBR Green PCR Master Mix和miScript Universal Primer,该引物对在逆转录反应中加入到cDNA的5'端的通用标签序列有特异性。单次cDNA合成反应足够用于许多miRNAs。使用miScript HiSpec Buffer制备的cDNA,配合miScript miRNA PCR Arrays或miScript Primer Assays可用于定量成熟miRNA。使用miScript HiFlex Buffer制备的cDNA配合real-time PCR步骤中合适的引物:用于miRNA或其它非编码RNA的miScript Primer Assays、用于miRNA前体的miScript Precursor Assays或用于mRNA的QuantiTect Primer Assays,可用于检测不同种类的RNA。 

Procedure

使用the miScript II RT Kit进行逆转录反应,得到的cDNA作为模板,配合miScript SYBR Green PCR Kit和合适的miScript Primer Assays和miScript miRNA PCR Arrays进行real-time PCR。使得miRNA定量和分析简单可行,实验室的real-time PCR循环仪上均可进行。

Applications

miScript SYBR Green PCR Kit作为miScript PCR System的一部分,用于:

  • 成熟miRNA的定量检测和谱分析
  • 同时检测miRNA和mRNA
  • 同时检测miRNA和miRNA前体
  • 检测snoRNA和其他非编码RNA

Supporting data and figures

Resources

Safety Data Sheets (1)

FAQ

What is the sequence of the miScript Universal Primer?
The sequence of the miScript Universal Primer in the miScript SYBR Green kit is proprietary.
FAQ ID -2994
Is the expression level of miRNA precursors lower compared to that of mature miRNAs?
In general, precursor miRNA (pre-miRNA) levels are lower than those of mature miRNAs. However this is not always the case. Sometimes the levels of both mature and pre-miRNA may be similar, or pre-miRNA levels may be higher. For this reason, researchers often want to quantify pre-miRNA as well as mature miRNA.  QIAGEN offers miScript precursor miRNA assays as well as mature assays for many miRNAs. 
FAQ ID -2807
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