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

S_2139_GEF_miRNY_small

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

miRNeasy FFPE Kit (50)

Cat. No. / ID:  217504

50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
Copy order details
$596.00
Log in To see your account pricing.
miRNeasy FFPE Kitは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 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

Product Details

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。

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”)。
See figures

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。
See figures

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。

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, qPRC, real-time RT-PCR, microArray
Elution volume14-30 µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinmiRNA, total RNA
Sample amountup 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)
ProcessingManual (protocol for automated processing on QIAcube available)
Main sample typeFFPE tissue samples
FormatSpin column
TechnologySilica technology
YieldVaries

Resources

パンフレット (7)
Sample to Insight solutions for successful molecular analysis
High-quality, nucleic acid purification for successful PCR and NGS experiments.
Critical factors for molecular analysis of FFPE samples
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
サイエンティフィック・ポスター (1)
Poster for download
クイックスタートプロトコール (2)
キットハンドブック (1)
追加リソース (1)
Safety Data Sheets (1)
Certificates of Analysis (1)
Brochures & Guides (6)
High-quality, nucleic acid purification for successful PCR and NGS experiments.
Sample to Insight solutions for successful molecular analysis
Critical factors for molecular analysis of FFPE samples
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
miRNA Research (1)
Kit Handbooks (1)
Additional Resources (1)
Scientific Posters (1)
Poster for download

FAQ

Are there specific recommendations for performing RT-PCR on RNA isolated from paraffin-embedded samples?

Paraffin-embedded or fixed samples typically yield fragmented, partially degraded RNA. In addition, RNA quality will depend greatly on the handling of the samples before, during, and after the fixation procedure. 

If performing RT-PCR with degraded RNA, we recommend using gene-specific primers or random nonamers rather than oligo-dT primers, since the mRNA poly-A tail may have been lost due to degradation.

We recommend that the RNeasy FFPE or miRNeasy FFPE kit be used to isolate the RNA.

FAQ ID -828
What is the composition of Buffer PKD?
The exact composition of Buffer PKD is proprietary. Buffer PKD functions as a Proteinase K Digestion Buffer and is a component of, for example, the AllPrep DNA/RNA FFPE Kit, RNeasy FFPE Kit, and the miRNeasy FFPE Kit. We are sometimes asked if Buffer PKD comprises any RNase inhibitors or RNase inhibiting agents — since the formalin-fixation of the starting material has already inactivated the RNases, no such reagents are present in this buffer.
FAQ ID -2801
I received a kit containing the MinElute columns; however, they were left out for a while and not stored at 2–8°C upon receipt. Can I still use them?

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560
What is the composition of Buffer RWT?
The exact composition of Buffer RWT is confidential. Buffer RWT is a proprietary component of, for example, the miRNeasy Mini Kit and the RNeasy Plus Universal Kit. Guanidine salt and ethanol are important ingredients in Buffer RWT. Ethanol is added by the user prior to the first use of the kit. Buffer RWT is a stringent washing buffer used after preclearing the sample with QIAzol Lysis Reagent, especially if isolation of small RNAs, for example, microRNAs or RNAs from formalin-fixed tissue, is desired
FAQ ID -2798
How do I clean up RNA preparations containing miRNA?

RNA preparations containing miRNA can be cleaned up by modifying* the cleanup protocols listed in the handbooks of the RNeasy Mini Kit or the RNeasy MinElute Cleanup Kit .

 

* Modify the cleanup protocol at step 2, by increasing the volume of ethanol (96-100%) from 250 µl to 950 µl.

FAQ ID -3002
Can small miRNA-containing RNA fractions be separated from large RNAs using the miRNeasy FFPE Kit?

Unlike the Appendix A Protocol for the miRNeasy Mini Kit, which allows removal of larger RNAs such as mRNA and rRNA to enrich miRNA in a separate small RNA fraction, this is not practical using the miRNeasy FFPE Kit. RNA from FFPE- or other compromised materials is usually strongly fragmented already. Significant amounts of rRNA and mRNA fragments would end up in the small RNA-enriched fraction.

 

FAQ ID -1737
What is the composition of Buffer RPE?
The exact composition of Buffer RPE is confidential. Buffer RPE is a mild washing buffer, and a proprietary component of RNeasy Kits. Its main function is to remove traces of salts, which are still on the column due to buffers used earlier in the protocol. Ethanol, which is added by the user just before using the kit for the first time, is an important ingredient of Buffer RPE.
FAQ ID -2797