miRNeasy FFPE Kit

ホルマリン固定パラフィン包埋した組織切片からのmicroRNAおよびトータルRNA精製
  • miRNA とトータル RNA の効率的な精製
  • ホルマリンによるクロスリンクを切断する新しい手法
  • RNA分解がなく効率的にRNAを遊離
  • わずか85 分でRNA が得られる能率化されたプロトコール

miRNeasy FFPE Kit は、ホルマリン固定パラフィン包埋(FFPE)組織切片から約18 ヌクレオチド(nt)以上のRNA を含むトータルRNA を精製します。本キットはリアルタイム定量RT-PCR のようなアプリケーションに利用できるmiRNA やその他のsmall RNA を含むRNA フラグメントを精製します。

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miRNeasy FFPE Kit (50)
50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
217504
kr. 4.015,00
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The miRNeasy FFPE Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Efficient RNA purification from FFPE tissues.|Superior yields and performance.|Effective real-time RT-PCR quantification.|miRNeasy FFPE procedure.|
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, CT 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, CT values were similar for both methods. CT values were similar or lower after purification using the miRNeasy FFPE Kit for all tissue types tested.|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 CT values 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.|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.||
Performance
miRNeasy FFPE Kitを用いたトータルRNAおよびmiRNAの収量と性能は、フェノール・クロロホルム抽出法や、他社の類似のキットを使用するmiRNA 精製法より優れています(図“効率的なリアルタイム定量RT-PCR”、FFPE組織からの効率的なRNA精製”および“卓越した収量および性能”)。
Principle

ホルマリン固定された組織では、ダウンストリームアプリケーションでRNA の性能を阻害するRNA・RNA およびRNA・タンパク質間のクロスリンキングが生じます。miRNeasy FFPE Kit は、特別な溶解およびインキュベーション条件によりRNA のホルマリンによるクロスリンキングを元に戻します。特別に開発された溶解バッファーは、組織切片からRNA を効率的に遊離し、さらにRNA 分解を防ぎます。最適な結合条件では、約18 ヌクレオチド以上のトータルRNA の精製を実現します。

Procedure
miRNeasy FFPE の全ての操作は、わずか85 分で完了します。至適化された溶解バッファーはRNA 収量を低減することなく、わずか15 分のProteinase K 分解でサンプル溶解が可能です。溶解後、サンプルを80 ℃で15 分間インキュベートし、ホルマリンによるクロスリンキングを元に戻します。その後、 DNase Booster Bufferとの至適化されたDNaseステップでゲノムDNAを迅速に除去し、RNeasy MinElute Spin Column を用いて高濃度なRNA を精製します(フローチャート“miRNeasy FFPE操作手順”)。RNA をわずか14 ~ 30 μl で溶出するため、より少量の反応容量でダウンストリームアプリケーションを行なえます。
Applications

miRNeasy FFPE Kit は、リアルタイム定量RT-PCR のような様々なアプリケーションで使用できるトータルRNA を含むmiRNA を精製します。

Feature
Specifications
Applications PCR, qPRC, real-time RT-PCR, microArray
Elution volume 14-30 µl
Format Spin column
Main sample type FFPE tissue samples
Processing Manual (protocol for automated processing on QIAcube available)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein miRNA, total RNA
Sample amount up to 4 sections, each with a thickness up to 10 µm and a surface area up to 250mm^2 (automated protocol on QIAcube: up to 2 sections)
Technology Silica technology
Yield Varies

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Kit Handbooks
1
For purification of total RNA, including miRNA, from formalin-fixed, paraffin-embedded tissue sections
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Scientific Posters
1
Images
Efficient RNA Purification from FFPE Tissues
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, CT 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, CT values were similar for both methods. CT values were similar or lower after purification using the miRNeasy FFPE Kit for all tissue types tested.
Superior Yields and Performance
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 CT values 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 of miRNA from FFPE tissues
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.
miRNeasy FFPE procedure
miRNeasy FFPE procedure.