miScript Primer Assays
配合miScript PCR System定量检测成熟miRNA
Features
- 灵敏并特异性的定量检测miRNA
- 对miRBase数据库中未列出的成熟miRNAs，提供用户订制服务
- 可在GeneGlobe便利的订购
Product Details
miScript Primer Assays是miRNA特异性引物，适合最新版本miRBase数据库中的所有人类、小鼠、大鼠和狗的miRNAs。miScript Primer Assays作为miScript PCR System的一部分，用于定量检测miRNA。cDNA使用miScript II RT Kit配合miScript HiSpec Buffer（只适用于成熟miRNA检测）或miScript HiFlex Buffer（使用相应的检测试剂同时定量检测各物种RNA）制备。cDNA作为模板，配合miScript Primer Assay和miScript SYBR Green PCR Kit进行real-time PCR。
Performance
miScript Primer Assays配合miScript PCR System检测成熟miRNA，结果可靠、重复性好。由于许多miRNAs表达水平低，灵敏度对数据的可靠性具有关键作用。miScript PCR System灵敏度高，可检测低至10拷贝的miRNA（参见" Accurate detection of as low as 10 miRNA copies"）。只需10 pg的总RNA，即可得到准确、重复性好的miRNA数据（参见" Broad dynamic range and high amplification efficiencies"）。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。
See figures
Principle
miScript PCR System配合SYBR® Green-based real-time PCR，灵敏和特异性定量检测和分析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。
使用miScript PCR System定量检测成熟miRNA，需要用到：
Procedure
成熟miRNA检测实验中，使用miScript II RT Kit中的miScript HiSpec Buffer或miScript HiFlex Buffer，通过逆转录反应制备cDNA，作为real-time PCR模板。使用miRNA特异性的miScript Primer Assay（正向引物）和包含miScript Universal Primer（反向引物）和QuantiTect SYBR Green PCR Master Mix的miScript SYBR Green PCR Kit，进行real-time PCR反应。
其它非编码RNA检测实验中，使用miScript II RT Kit中的miScript HiFlex Buffer，通过逆转录反应制备cDNA，作为real-time PCR模板。使用miScript Primer Assay（正向引物）和包含miScript Universal Primer（反向引物）和QuantiTect SYBR Green PCR Master Mix的miScript SYBR Green PCR Kit，进行real-time PCR反应。如果没有针对目的小RNA的miScript Primer Assay，可以订制设计。
Applications
miScript Primer Assays作为miScript PCR System的一部分，用于：
- 定量检测成熟miRNA
- snoRNA检测
Supporting data and figures
Accurate detection of as low as 10 miRNA copies.
Synthetic miR-21 was used to generate cDNA using the miScript II RT Kit with [A] miScript HiSpec Buffer or [B] miScript HiFlex Buffer (both provided in the kit). A range of amounts from 10 copies to 106 copies of this cDNA was used in real-time PCR using the miScript PCR System. Real-time PCR was performed on the Rotor-Gene Q cycler. The resultant CT values decreased linearly with increasing miRNA copy number, indicating sensitive detection from a wide range of template amounts.
Resources
Safety Data Sheets (1)
FAQ
3322 - What is the concentration of the reconstituted miScript Primer Assay (100) and miScript Precursor Assay (100) tubes?
Are miScript Primer Assay included in miScript miRNA PCR Arrays the same primers as the miScript Primer Assays available in individual tubes?
Is the expression level of miRNA precursors lower compared to that of mature miRNAs?
Are miScript Primer Assays available for plant mature miRNAs?
How reliable are the results from the miScript Primer Assays?
Are plant miScript Primer Assay designs wet-bench validated?