miScript miRNA PCR Arrays

使用miScript PCR System对miRNA进行基于SYBR® Green法的real-time PCR分析

Products

miScript miRNA PCR Arrays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。
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miRNome V16

Cat. No. / ID:  331222

miRNome panels of miRNA assays in 96-well, 384-well, or Rotor-Disc formats
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miScript miRNA HC PCR Array

Cat. No. / ID:  331223

High-content array of assays for a pathway, disease, or gene family miRNAs; available in 384-well format
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miScript miRNA QC PCR Array

Cat. No. / ID:  331292

Array of quality control assays available in 96-well, 384-well, or Rotor-Disc format
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Pathway-Focused miScript miRNA PCR Array

Cat. No. / ID:  331221

Pathway- or disease-focused panels of miRNA assays in 96-well, 384-well, or Rotor-Disc formats
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Features

  • 灵敏和特异性的进行miRNA谱分析
  • 分析信号通路和疾病相关的miRNAs或全miRNA组
  • 免费提供在线工具,方便进行数据分析

Product Details

miScript miRNA PCR Arrays是成熟miRNA的特异性正向引物(miScript Primer Assays),可对生物相关的信号通路和全miRNA组平台进行分析。作为miScript PCR System的一部分,miScript miRNA PCR Arrays用于miRNA定量检测。miScript II RT Kit配合miScript HiSpec Buffer得到的cDNA作为real-time PCR的模板,配合miScript miRNA PCR Array和miScript SYBR Green PCR Kit使用。

Performance

发现血清或血浆样本中的生物标记

以信号通路为主的miScript miRNA PCR Arrays的检测范围不断扩大,可发现生物过程中miRNAs的作用。使用QIAGEN专有的方法,可选择miScript miRNA PCR Arrays的量,确保芯片时时更新且与生物研究息息相关。目前miRNA研究领域突出的是血清或血浆样本中的miRNAs检测。近期发现血清和血浆中的胞外miRNAs相对稳定,研究集中在miRNA水平的变化,可作为一系列疾病研究的潜在非侵入性生物标记。设计Serum & Plasma miScript miRNA PCR Array,可快速分析84个与血清和血浆最具相关性的miRNAs(参见" Analysis of expression of circulating miRNAs in serum or plasma")。

单个cDNA样本完成所有miRNomes分析

除了用于信号通路研究的miScript miRNA PCR Arrays,还有及时更新的miRNome miScript miRNA PCR Arrays用于包括人类、小鼠、大鼠和狗等多个物种。使用不到1 µg RNA即可快速分析人类的所有miRNome(参见" Whole miRNome profiling")。miScript miRNA PCR Array中的每个引物都是经过实验验证的,确保使用real-time PCR高灵敏度、高特异性检测成熟miRNA。

See figures

Principle

miScript PCR System配合SYBR Green-based real-time PCR,高灵敏度、高特异性进行miRNA定量检测和分析。可以完成从RNA中合成cDNA及后续的real-time PCR检测miRNAs涉及的所有步骤。

使用miScript PCR System分析成熟miRNA,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。
  • miScript miRNA PCR Array,前沿的miScript miRNA PCR Arrays包含成熟miRNA的特异性正向引物(miScript Primer Assays),用于miRNome平台及生物相关的、以信号通路为主的平台。

Procedure

使用miScript miRNA PCR Arrays进行miRNA表达分析简单、可靠。首先,使用miScript HiSpec Buffer配合miScript II RT Kit制备cDNA。然后,加入cDNA预混物、miScript Universal Primer、QuantiTect SYBR Green PCR Master Mix和不含RNase的纯水到miScript miRNA PCR Array。接着,运行real-time PCR循环仪,进行反应。最后,使用miScript miRNA PCR Array Data Analysis Tool分析数据。
miScript miRNA PCR Array板布局

miScript miRNA PCR Arrays适用于96孔板、384孔板或Rotor-Disc 100孔盘三种规格(参见" Pathway-Focused or miRNome miScript miRNA PCR Array layout for plate formats A, C, D, F"、" Pathway-Focused miScript miRNA PCR Array layout for plate formats E, G"、" 384HC and miRNome miScript miRNA PCR Array layout for plate formats E, G"和" miScript miRNA PCR Array layout for Rotor-Disc format R")。 每个芯片都包含几个对照检测,检测目的见表。

miScript miRNA PCR Arrays对照检测
对照 目的
C. elegans miR-39 miScript Primer
Assay
使用外源标准Syn-cel-miR-39 miScript miRNA Mimic,得到可供选择的标准化数据
6 snoRNA/snRNA miScript Primer
Assays (miScript PCR Controls)
使用相对定量的ΔΔCT方法得到标准化数据
miRNA reverse transcription control
(miRTC)
评估逆转录性能
Positive PCR control (PPC) 评估PCR性能

有关上述对照的更多信息,参见miScript miRNA PCR Array Handbook

miScript miRNA QC PCR Array板布局

miScript miRNA QC PCR Arrays使用real-time PCR进行多个cDNA样本的质量评估。提供96孔板、384孔板或Rotor-Disc 100三种规格(如需更多信息,参见miScript miRNA PCR Array Handbook)。每个miScript miRNA QC PCR Array包含在miScript miRNA PCR Array上进行对照检测的副孔。96孔板和Rotor-Disc 100规格为至多8个cDNA样本提供质量对照。384孔板规格为至多32个cDNA样本提供质量对照。

miScript miRNA PCR Array Data Analysis Tool

在线提供关于miScript miRNA PCR Arrays的即用型数据分析软件,操作方便。每个miScript miRNA PCR Array带有一个专有的数据分析工具。一旦输入原始CT值,该分析工具即可自动分析PCR对照,使用∆∆CT方法进行相对定量。通过多种格式显示结果,可快速解读数据。

See figures

Applications

miScript miRNA PCR Arrays是miScript PCR System产品线的一部分,适用于:

  • 分析疾病或信号通路相关的miRNA
  • 全miRNA组分析
  • 生物标记物发现

Supporting data and figures

Resources

Safety Data Sheets (6)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
产品介绍与指南 (4)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
安全数据表 (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (3)

miScript II RT Kit

miScript SYBR Green PCR Kit

miScript miRNA PCR Arrays

miScript miRNA QC PCR Array

For SYBR Green-based, real-time PCR profiling of microRNAs using pathway-focused arrays, HC arrays, and miRNome arrays

仪器技术参数 (2)
For gene expression and genomic analysis
For pathway-focused miRNA or miRNome analysis
下载文件 (1)
Data analysis file for miScript miRNA PCR Array All miRNA QC (96-well plate, Rotor-Disc 100)
Catalog number- 331221
Pathway number- MIXX-989Z
Kit Handbooks (3)

miScript II RT Kit

miScript SYBR Green PCR Kit

miScript miRNA PCR Arrays

miScript miRNA QC PCR Array

For SYBR Green-based, real-time PCR profiling of microRNAs using pathway-focused arrays, HC arrays, and miRNome arrays

Certificates of Analysis (1)
Brochures & Guides (4)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
Download Files (1)
Data analysis file for miScript miRNA PCR Array All miRNA QC (96-well plate, Rotor-Disc 100)
Catalog number- 331221
Pathway number- MIXX-989Z
Instrument Technical Documents (2)
For pathway-focused miRNA or miRNome analysis
For gene expression and genomic analysis
miRNA Research (1)

FAQ

Can miScript miRNA qPCR Assays detect single nucleotide difference between miRNAs?
Yes. With our patent pending proprietary technology, miScript miRNA qPCR Arrays and Assays can distinguish miRNAs that differ by only one single nucleotide.
FAQ ID -2723
Will pipetting error affect the miRNA qPCR Assay results?
The passive reference dye, ROX in the miScript PCR master mix is used by real-time PCR systems to normalize variation from well to well. Therefore, these systems tolerate volume variations caused by pipetting error and evaporation.
FAQ ID -2728
Do I need the sequence specific miRNA primers in the RT step of the miRNA qPCR Assays?
No. Because the miScript miRNA qPCR assay technology uses universal reverse transcription in the RT step, sequence specific miRNA primers are not needed in this step. Therefore, cDNA obtained from universal RT can be used for any miScript miRNA expression analysis qPCR assay or PCR array.
FAQ ID -2725
How can I prevent the evaporation of reaction volume from the wells in the miScript miRNA PCR array?
Be sure to carefully and completely seal the PCR plate with the optical thin-wall 8-cap strips or the optical adhesive film before placing it into your thermal cycler. Also, be sure to use a compression pad when using an optical film seal as directed by the manufacturer of your real-time PCR instrument.
FAQ ID -2729
Why did the real-time PCR yield Ct values < 12?
You probably used too much template. Try less input RNA for reverse transcription especially if the higher end of the recommended range had been used previously. Or use a smaller volume of template per PCR reaction or use diluted template, but do not use less than 1 µl of template per each 25-µl miRNA PCR array reaction. Remember to use the same volume of template in each reaction.
FAQ ID -2727
Can miScript miRNA qPCR Assays be used for pre-miRNA detection?
No. miScript miRNA PCR Arrays and miScript Primer Assays are designed to measure exclusively the expression of mature miRNA, not pre-miRNA. To measure miRNA precursors use miScript Precursor Assays available through the QIAGEN GeneGlobe assay search portal.
FAQ ID -2724
Can I use total RNA for the miRNA PCR Arrays or Assays?
Yes, you can. In fact, total RNA is the recommended starting material for the miScript PCR System. We recommend using the miRNeasy Mini Kit (217004) to isolate total RNA for use with the miScript PCR System.
FAQ ID -2726