配合miScript PCR System定量检测miRNA前体

  • 灵敏和特异性的定量检测miRNA前体
  • 同时检测前体和成熟miRNA
  • 可在GeneGlobe便利的订购

miScript Precursor Assays是miRNA前体的特异性正向和反向引物,配合miScript SYBR Green PCR Kit进行miRNA前体的real-time PCR。适合最新版本miRBase中的所有人类、小鼠和大鼠miRNAs,miRBase中的数据会不断更新。miScript II RT Kit可配合miScript HiFlex Buffer制备cDNA。cDNA作为模板,配合miScript Precursor Assay和miScript SYBR Green PCR Kit中的QuantiTect SYBR Green PCR Master Mix用于real-time PCR。

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You can search for the following terms:
  • Entrez Gene IDs (e.g., 835)
  • RefSeq IDs (e.g., NM_032983, NP_116765)
  • Gene symbols (e.g., CASP2)
  • Cat. no. (e.g., SI00299551, QT01342509)
  • Sanger ID or Accession (e.g., hsa-let-7b, MI0000063)
  • CpG loci identification numbers (CG#) (e.g., CG17753661)

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miScript Precursor Assay (100)
10x miScript Precursor Assay (contains one miRNA-precursor–specific forward primer and one miRNA-precursor–specific reverse primer)
218600 Varies
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miScript Precursor Assays are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


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Parallel detection of mature and precursor miRNA.|
Total RNA was purified from HeLa S3 cells using the miRNeasy Mini Kit. cDNA was prepared using the miScript II RT Kit with the provided miScript HiFlex Buffer. In real-time PCR, miScript Primer Assays were used for mature miRNA quantification and miScript Precursor Assays were used for precursor miRNA quantification. The miScript PCR Control for RNU6-2 was used for normalization. Real-time PCR was carried out on the Rotor-Gene Q cycler. Using the miScript PCR System, mature miRNAs and their precursor miRNAs can be efficiently quantified in parallel real-time PCR reactions.|
Performance

miScript Precursor Assays可准确检测miRNAs前体,可用于需同时检测miRNAs前体和成熟miRNAs的实验(参见"Parallel detection of mature and precursor miRNA")。

Principle

miScript PCR System配合SYBR® Green-based real-time PCR,灵敏和特异性地检测并定量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进行miRNA前体检测和定量,需要以下组件:

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。如需检测miRNA前体,无需使用miScript Universal Primer。
miScript Precursor Assay,包含miRNA前体特异性正向和反向引物,配合miRBase中的最新信息,检测miRNA前体。miRNA前体检测可与成熟miRNA(使用miScript Primer Assay检测)和mRNA(使用QuantiTect Primer Assay检测)检测同时进行,使用相同的cDNA样本。miScript Universal Primer不用于检测miRNA前体。
miRNA前体研究

miScript Precursor Assays包含miRNA前体特异性正向和反向引物,靶向作用于研究中的miRNA前体的茎环序列。miScript Precursor Assay靶向于初级miRNA(pri-miRNA)和miRNA前体的茎环序列,因此miScript Precursor Assays可同时定量初级miRNA和miRNA前体。miRNA前体检测可用于miRNA生物研究、调控和功能研究。不同miRNAs的成熟和miRNA前体水平各有不同。另外,在不同条件和组织中,成熟miRNAs可从不同的miRNAs前体中表达。

Procedure

miRNA前体检测实验中,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的模板。Real-time PCR配合miScript Precursor Assay和miScript SYBR Green PCR Kit中的QuantiTect SYBR Green PCR Master Mix进行。

Applications

miScript Precursor Assays作为miScript PCR System的一部分,用于miRNA前体的检测。

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Kit Handbooks
1
For real-time PCR analysis of microRNA using SYBR Green detection
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