RNeasy FFPE Kit



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RNeasy FFPE Kit (50)

Cat. No. / ID:  73504

50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
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RNeasy 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


  • 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

RNeasy FFPE Kitはホルマリン固定、パラフィン包埋組織切片からのトータルRNA精製用に特別にデザインされています。特別な溶解およびインキュベーション条件によりホルムアルデヒドによるクロスリンクを元に戻します。さらに溶解バッファーは、組織切片からRNAを効率的に遊離し、操作におけるRNA分解もありません。また本キットでは、混入したゲノムDNAコンタミを最適に除去するためにDNaseとDNase Booster Bufferを使用しています。また、わずか14 μl の溶出容量でのトータルRNA 精製が可能なRNeasy MinElute スピンカラムを使用しています。精製はQIAcube上で全自動化できます。


RNeasy FFPE Kitを用いることにより、固定および包埋サンプルから精製されるRNA収量は、他の精製法を用いたものよりも高くなります(図" RNA収量の増加")。70ヌクレオチドより長い全てのRNAが回収され、RNAの分解、ゲノムDNAのコンタミもほとんど認められません(図" 全ての使用可能なRNAの回収")。本キットで精製したRNA は、リアルタイムRT-PCRなどの様々なアプリケーションで最適な結果を実現します(図" リアルタイムRT-PCR解析における信頼性の高い結果"および" リアルタイムRT-PCR解析の成功")。
See figures


ホルマリン固定された組織はRNA–RNAおよびRNA–タンパク質間のクロスリンクが生じているため、酵素によるアッセイ系でのRNAの使用において良好な結果が得られません。FFPEサンプルでは、固定および包埋条件も核酸の過度なフラグメント化をもたらします。RNeasy 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ホルムアルデヒドによるクロスリンクを元に戻します。溶解バッファーはFFPE組織サンプルからRNAを効率的に遊離させ、RNA分解を回避します。これらの至適条件により、全ての使用可能なRNAの精製が可能になるため、RNeasy FFPE KitでFFPEサンプルから精製されるRNA収量は、ほかの精製法を用いたものよりも収量よりも大きくなります。


RNeasy FFPEの全ての操作手順は、わずか85分で完了します(フローチャート"RNeasy FFPE操作手順")。proteinase K分解によるサンプル溶解はわずか15分で完了します。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クを元に戻します。次に、DNaseを用いて、小さなDNAフラグメントを含むゲノムDNAを効率的に除去します。最後に、RNeasy MinElute Spin Columnを用いてRNAを14〜30μlで溶出し、高濃度なRNAを精製します。精製工程はQIAcube上で全自動化できます。
See figures


RNeasy FFPE Kitは、70 ヌクレオチドより長い全てのRNAを分離するので、RT-PCRなどの多数のアプリケーションに有用なRNAフラグメントを回収できます。しかし、FFPEのサンプルから精製されたRNAはフラグメント化が激しく、完全長RNAを必要とするダウンストリームアプリケーションには使用するべきではありません。いくつかのアプリケーションではフラグメント化したRNAを使用できるように変更が必要な場合があります(例:RT-PCRで短い増幅産物にするようにプライマーをデザインする)。cDNA合成にはオリゴ-dTプライマーの代わりにランダムプライマーか遺伝子特異的なプライマーを使用するべきです。

Supporting data and figures


ApplicationsPCR, qPCR, real-time RT-PCR, microarray
Elution volume14–30 µl
Main sample typeFFPE tissue samples
ProcessingManual (centrifugation), automated (QIAcube)
FormatSpin column
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinRNA
Time per run or per prep70 minutes
Sample amount1*5 µm to 4*10µm sections
TechnologySilica technology


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.
FFPE サンプルの分子解析における重要なファクター
Critical factors for molecular analysis of FFPE samples
MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
キットハンドブック (1)
追加リソース (1)
クイックスタートプロトコール (1)
For purification of total RNA from formalin-fixed, paraffin-embedded tissue sections
Technical Information and Important Notes (1)
September 2023
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
FFPE サンプルの分子解析における重要なファクター
Gene Expression Analysis (1)
Additional Resources (1)
Quick-Start Protocols (1)
For purification of total RNA from formalin-fixed, paraffin-embedded tissue sections
Kit Handbooks (1)


Determinants of RNA quality from FFPE samples.
von Ahlfen S; Missel A; Bendrat K; Schlumpberger M;
PLoS One; 2007; 2 (12):e1261 2007 Dec 5 PMID:18060057


Do you have a kit for the isolation of RNA from formalin-fixed, paraffin-embedded samples?

Yes, we offer the RNeasy FFPE Kit specially designed to purify total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections.  It provides special lysis and incubation conditions to reverse formaldehyde modification of RNA typical for formalin fixed tissues.

FAQ ID -1174
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
Do you have a kit for RNA isolation from any kind of sample type?

The RNeasy 96 Universal Tissue Kit enables high-throughput purification of RNA from any animal or human tissue sample, including difficult-to-lyse fibrous and fatty tissues. For single tube format RNA purification, the RNeasy Plus Universal Kit is also available. 

Please refer to the Selection guide for RNA isolation for all sample types to find the optimal solution for your sample source.

FAQ ID -627
What is the composition of Buffer RLT?

The exact composition of Buffer RLT is confidential. This buffer is a proprietary component of RNeasy Kits. Buffer RLT contains a high concentration of guanidine isothiocycanate, which supports the binding of RNA to the silica membrane. Buffer RLT can be purchased separately (cat. no. 79216)

Note: note that ß-mercaptoethanol should be added to Buffer RLT before use to effectively inactivate RNAses in the lysate (10 µl ß-Mercaptoethanol per 1 ml Buffer RLT).

FAQ ID -2793
How can I check the integrity of RNA purified using RNeasy Kits?

The integrity and size distribution of total RNA purified with RNeasy Kits can be checked by denaturing-agarose gel electrophoresis, the Agilent 2100 bioanalyzer, or the QIAxcel Advanced System with the QIAxcel RNA QC Kit v2.0.

The respective ribosomal species should appear as sharp bands on the stained gel. 28S ribosomal RNA bands should be present with an intensity approximately twice that of the 18S RNA band. If the ribosomal bands are not sharp, but appear as a smear of smaller sized RNAs, it is likely that the RNA sample has suffered major degradation during preparation.

Size of ribosomal RNAs from various sources



Size (kb)

E. coli






S. cerevisiae



























FAQ ID -1024
What is the maximum volume of RNA in solution that can be used with the QuantiTect Whole Transcriptome Kit?

A maximum of 5 µl RNA eluate from RNeasy extraction procedures can be added to the reverse-transcription reaction with the QuantiTect Whole Transcriptome Kit.



FAQ ID -1616
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
What is Tissue-Tek O.C.T., and what is it used for?

Tissue-Tek O.C.T. is an embedding compound for cryosectioning, which is soluble in water. It mainly consists of glycols and synthetic resins. Tissue-Tek O.C.T. is used as matrix for cryosectioning of tissues. Using the O.C.T. the tissue samples can be positioned more easily in the microtome and have better qualities during sectioning.

Most cryosections are fixed using non-crosslinking agents. For isolation of RNA from tissue embedded in Tissue-Tek O.C.T. using non-crosslinking agents the RNeasy Plus Micro Kit or the RNeasy Micro Kit (Protocol: Total RNA Isolation from Microdissected Cryosections), or the RNeasy Mini Kit (RNeasy Mini Protocol for the isolation of Total RNA from Animal Tissue) give great results. We strongly recommend removing as much of the embedding compound as possible prior to RNA extraction from the sections. Please see QIAGEN News article, Issue 1 1998, 'Effects of malnutrition on expression of lactase in children' for successful RNA isolation from O.C.T.-embedded tissue using the RNeasy Mini Kit.

In case crosslinking agents (e.g. formaldehyde or glyoxal-containing) were used for fixation of the tissue for cryosectioning the RNeasy FFPE Kit is the perfect choice. The RNeasy FFPE Kit is especially designed for purifying total RNA from formalin-fixed tissue sections. Special lysis and incubation conditions reverse formaldehyde modification of RNA for improved results in downstream application. The crosslinking causes the RNA to break, resulting in overall smaller molecules, which look like a smear when analyzed on a formaldehyde gel.

In case DNA as well as RNA from precious samples is of interest from the identical sections, please have a look at the AllPrep DNA/RNA Kits for unfixed tissue or the AllPrep DNA/RNA FFPE Kit for fixed tissue.

FAQ ID -530
What are the effects of low A260/A230 ratios in RNA preparations on downstream applications?

The efficiency of downstream applications depends strongly on the purity of the RNA sample used.  Pure RNA should yield an A260/A230 ratio of around 2 or slightly above; however, there is no consensus on the acceptable lower limit of this ratio.  Possible candidates that can increase the A230 include “salt”, carbohydrates, peptides, and phenol (or aromatic compounds in general).  In our experience, the increased absorbance at 230 nm in RNA samples is almost always due to contamination with guanidine thiocyanate, present at very high concentrations in the lysis buffer or extraction reagent used in most RNA purification procedures.

Please find an article discussing the effect of low 260/230 ratios in RNA preparations on downstream applications on page 7 of QIAGEN Newsletter March 15, 2010 . In summary, we found that concentrations of guanidine thiocyanate of up to 100 mM in an RNA sample do not compromise the reliability of downstream applications.



FAQ ID -2248
How can I ensure complete genomic DNA removal when using the RNase-Free DNase Set?

To ensure efficient gDNA removal when doing an on-column digest using the RNase-Free DNase Set in combination with RNeasy Mini the following factors are crucial:

  • prevent overloading by adjusting the amount of starting material to no more than the maximum amounts recommended in the RNeasy Mini Handbook
  • ensure complete disruption and homogenization of the starting material as instructed in the section 'Disruption and homogenization of starting materials' of the handbook
  • strictly follow the protocol for on-column DNase Digestion in Appendix D of the RNeasy Mini Handbook (you can let wash buffer RW1 incubate on the column for 3-5 minutes before centrifuging to enhance removal of excess gDNA prior to applying the enzyme)

In the rare case that trace amounts of genomic DNA are still detectable in sensitive downstream applications such as e.g., realtime RT-PCR, an in-solution digest using the RNase-Free DNAase set can be performed. Instructions are presented in Appendix C of the RNeasy MinElute Cleanup Handbook.

Alternatively, a second on-column digest can be carried out in future preparations, immediately following the RW1 wash after the first incubation with DNase.

FAQ ID -1087
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
What is the composition of Buffer RW1?

The exact composition of Buffer RW1 is confidential. Buffer RW1 is a proprietary component of RNeasy Kits. Buffer RW1 contains a guanidine salt, as well as ethanol, and is used as a stringent washing buffer that efficiently removes biomolecules such as carbohydrates, proteins, fatty acids etc., that are non-specifically bound to the silica membrane. At the same time, RNA molecules larger than 200 bases remain bound to the column.

Note: Buffer RW1 should not be used for isolation of small RNAs, for example, microRNAs or fragmented RNA from formalin-fixed tissues, as these smaller fragments will be washed away.  Buffer RWT should be used instead.

FAQ ID -2796