QIAGEN solutions for advancing microRNA research


miRNeasy Mini Kit and miRNeasy 96 Kit    
高效纯化含miRNA的总RNA或单独富集miRNA
  • 适合各种细胞和动物组织，处理其他样品（如FFPE组织、细菌）
        的方法即将推出，
  • 纯化>18 nt至200 nt的小RNA，可单独富集miRNA组分
  • 纯度高，可用于northern、real-time RT-PCR、microarray等分析
  • 选择灵活，提供离心柱和96孔板纯化方式
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    图1 高纯度RNA纯化，无苯酚污染
     
    图2 从不同起始量的组织中同时纯化miRNA和mRNA
    Total RNA including miRNA was purified from 25 mg of a range of RNAlater® stabilized rat tissues using the miRNeasy 96 Kit. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16 and for the larger mRNA of the PGK1 gene. Results showed successful detection of both PGK1 mRNA (large RNA) and miR-16 (small RNA) from the same eluates.
    		 

    图3 miRNeasy Kit纯化的结果优于TRIzol
         Real-time RT-PCR assays for the miRNA miR-16. Results showed that CT values were lower after purification using the miRNeasy Kit, indicating that higher amounts of miRNA were purified than when using TRIzol.



    The miScript System    		    
    使用SYBR Green based real-time PCR检测各种miRNA, 包含引物miScript Primer Assay、逆转录试剂miScript everse Transcription Kit、定量检测试剂miScript SYBR Green PCR Kit 三个部分，可单独购买。
  • 简单、快速地从同一cDNA中检测多种miRNA
  • 同一cDNA可用于同时检测miRNA、小RNA和mRNA
  • 特异性高，灵敏度高，起始样品低至10pg总RNA
  • GeneGlobe 数据库提供人、小鼠、大鼠miRNA特异性引物


    		•    
    The miScript System ——
    miScript Primer Assay
    提供miRNA数据库miRBase version 9.0中>95%的人、小鼠、大鼠miRNA对应的正向引物。
  • 通过www.qiagen.com/GeneGlobe 检索引物，方便快速
  • 人的primer已经通过实验验证
  • 选择灵活，提供单管装或96孔板set


    		•     
    The miScript System ——
    miScript Reverse Transcription Kit
    包含逆转录步骤所需全套试剂，以含miRNA的总RNA或单独的
    miRNA组分为转录模板，通过末端加A、通用引物完成逆转录。
  • miRNA、小RNA和mRNA同时逆转录得到cDNA
  • 线性范围广，从10pg到1ug总RNA，大于7个数量级
  • 操作简单，只需要两步孵育即可完成


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    图4单次cDNA合成反应可用于检测多种iRNAs和mRNAs
    Total RNA was prepared from HeLa S3 cells using the miRNeasy Mini Kit. The miScript System was used for real-time PCR analysis of 4 miRNAs (Let-7a, miR-16, miR-21, and miR-25). QuantiTect Primer Assays were used for real-time PCR analysis of 4 mRNAs (GAPDH, PPIA, CDC2, and Erk2).

    图5 高度线性的cDNA合成反应
    RNA was purified from HeLa S3 cells using the miRNeasy Mini Kit. A range of amounts of RNA from 10 pg to 1 μg were used in cDNA synthesis reactions using the miScript Reverse Transcription Kit. cDNA was used as a template in quantitative, real-time PCR assays for 5 miRNAs (miR-16, miR-20a, miR-25, miR-28, and miR-26a).
    The miScript System ——
    miScript SYBR Green PCR Kit
    使用SYBR Green real-time PCR方法定量检测miRNA, 以master mix预混液形式提供，含通用引物，只需另加正向引物即可。
  • 可用于检测miRNA、小RNA和mRNA，只需加入各自的引物即可
  • 灵敏度高，线性范围广，低至10pg总RNA
  • 特异性高，miRNA异构体也可区分（如Let-7 isoforms）
  • 预混液形式提供，操作简单、快速
    		•  

    Synthetic miRNAs of each Let-7 isoform were used in cDNA synthesis reactions performed with the miScript Reverse Transcription Kit. An aliquot of the resultant cDNA was used as a template in real-time PCR reactions with a miScript Primer Assay for each isoform and the miScript SYBR Green PCR Kit. The % relative detection was calculated using the differences between the CT values actieved from the mismatching miScript Primer Assays and those from the perfectly matching miScript Primer Assays (% relative detection = 2– ΔCT x 100).

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    		 W W W . Q I A G E N . C O M Advancing microRNA research
    由QIAGEN公司委托 生物通 制作