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Qiagen miRNeasy FFPE kit（50）

产品中心 >> 试剂 >> 分子生物学试剂 >> qPCR kits & 核酸抽提

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miRNeasy FFPE kit（50）

分类：qPCR kits & 核酸抽提

品牌：Qiagen

货号：217504

原价：￥5470

现价：￥4207.50 节省：￥1262

规格：50Tests

产品参数

从福尔马林固定、石蜡包埋的组织切片中进行miRNA和总RNA的纯化

有效纯化miRNA和总RNA
创新方法用于克服福尔马林造成的交联
有效释放RNA而不会破坏其完整性
85分钟流畅的操作得到RNA

miRNeasy FFPE Kit确保从福尔马林固定、石蜡包埋（FFPE）的组织切片中纯化含约18个核苷酸以上的总RNA。试剂盒能够回收可用的RNA片段，包括miRNA和其他小RNA，用于定量real-time RT-PCR等下游应用。

miRNeasy FFPE Kit 点此下载

For purification of microRNA and total RNA from formalin-fixed, paraffin-embedded tissue sections

Effective purification of miRNA and total RNA
Novel method to overcome formalin crosslinking
Efficient release of RNA without compromising integrity
Streamlined protocol providing RNA in just 85 minutes

The miRNeasy FFPE Kit enables purification of total RNA that includes RNA from approximately 18 nucleotides upwards from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The kit provides recovery of usable RNA fragments, including miRNA and other small RNAs, for applications such as quantitative, real-time RT-PCR.

Product Details

Performance

Yields and performance of total RNA and miRNA with the miRNeasy FFPE Kit are superior to alternative methods of miRNA purification, such as using phenol-chloroform extraction or a similar kit from another supplier (see figuresEffective real-time RT-PCR quantification, Efficient RNA purification from FFPE tissues and Superior yields and performance).

Principle

Fixing tissues with formalin leads to RNA-RNA and RNA-protein crosslinking which impairs RNA performance in downstream applications. The miRNeasy FFPE Kit provides special lysis and incubation conditions to reverse formalin crosslinking of RNA. A specially developed lysis buffer efficiently releases RNA from tissue sections while avoiding further RNA degradation. The lysate is treated with DNase then optimized binding conditions allow purification of all usable RNA from approximately 18 nucleotides upwards.

Procedure

The entire miRNeasy FFPE procedure can be completed in as little as 85 minutes. The optimized lysis buffer allows sample lysis with proteinase K digestion in only 15 minutes without sacrificing RNA yields. After lysis, samples are incubated at 80ºC for 15 minutes to reverse formalin crosslinking. Genomic DNA is then rapidly removed in an optimized DNase step with DNase Booster Buffer, and concentrated RNA is purified using RNeasy MinElute spin columns (see flowchart miRNeasy FFPE procedure). Since RNA is eluted in a volume of just 14–30 µl, smaller reaction volumes are possible in downstream applications.

Applications

The miRNeasy FFPE Kit allows purification of miRNA with total RNA for use in a variety of applications, such as quantitative, real-time RT-PCR.

Images

Efficient RNA purification from FFPE tissues.

Total RNA including miRNA was purified from the indicated rat tissues using either the miRNeasy FFPE Kit or phenol-chloroform extraction. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16 using the miScript PCR System. Results showed that for lung and liver tissues, C_T values were lower after purification using the miRNeasy FFPE Kit, indicating that higher amounts of miRNA were purified than when using phenol-chloroform extraction. For kidney tissue, C_T values were similar for both methods. C_T values were similar or lower after purification using the miRNeasy FFPE Kit for all tissue types tested.

Superior yields and performance.

RNA was purified from FFPE sections of the indicated rat tissues using either the miRNeasy FFPE Kit or a similar kit from Supplier A. [A] RNA yields were determined by measuring absorbance at 260 nm. [B] Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16 using the miScript PCR System. Results showed that C_Tvalues were lower after purification using the miRNeasy FFPE Kit, indicating that higher amounts of miRNA were purified than when using the kit from Supplier A.

Effective real-time RT-PCR quantification.

Rat liver tissue was formalin fixed for 24 hours or 60 hours, followed by purification of total RNA including miRNA using the miRNeasy FFPE Kit. Purified RNA was used as a template in quantitative, real-time RT-PCR using the miScript PCR System to detect miRNAs miR-16 and miR-29a and for the larger mRNA of the PGK1 gene. Results show successful detection of both miRNAs as well as for the mRNA from the same eluate.