miScript SYBR® Green PCR Kit
配合miScript PCR System,使用real-time PCR方法检测miRNA
配合miScript PCR System,使用real-time PCR方法检测miRNA
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。
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 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。
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。
miScript SYBR Green PCR Kit作为miScript PCR System的一部分,用于:
cDNA was prepared from human colon adenocarcinoma total RNA and normal adjacent colon total RNA. miRNome profiling was performed using the Human miRNome miScript miRNA PCR Array. Fifty-six miRNAs were strongly downregulated and 50 were strongly upregulated in the adenocarcinoma tissue (red lines indicate ±4-fold change).