miRNeasy FFPE Kit

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

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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

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miRNeasy FFPE Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

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

Product Details

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。

Performance

使用miRNeasy FFPE Kit纯化的总RNA和miRNA的产量和性能优于采用其他方法纯化的miRNA，如使用苯酚-氯仿抽提或者其他供应商的类似试剂盒（参见""、""和""）。

See figures

Effective real-time RT-PCR quantification.

Efficient RNA purification from FFPE tissues.

Superior yields and performance.

Principle

用福尔马林固定的组织导致RNA-RNA和RNA-蛋白质交联，从而影响了RNA在下游应用中的表现。miRNeasy FFPE Kit提供了独特的裂解和孵育条件，可逆转福尔马林对RNA造成的交联。特别研制的的裂解缓冲液有效地将RNA从组织切片中释放出来，同时避免了RNA的进一步降解。裂解产物用DNase消化，然后优化的结合条件能够纯化含约18个以上核苷酸的所有可用的RNA。

Procedure

所有的miRNeasy FFPE操作流程都可在85分钟之内完成。优化的裂解缓冲液能够用蛋白质酶K消化样本裂解产物，仅需15分钟而且不损耗RNA产量。裂解后，在80°C下孵育样本15分钟逆转福尔马林交联。然后，在优化的DNase消化步骤中，用DNase Booster Buffer快速去除基因组DNA，并且用RNeasy MinElute离心柱纯化浓缩的RNA（参见""）。由于RNA的洗脱体积仅有14–30 µl，在下游应用中可实现较小的反应体积。

See figures

miRNeasy FFPE procedure.

Applications

miRNeasy FFPE Kit能够纯化包含miRNA的总RNA，并且适用于多种下游应用，如定量real-time RT-PCR。

Supporting data and figures

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. 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.

Specifications

Features	Specifications
Applications	PCR, qPRC, real-time RT-PCR, microArray
Elution volume	14-30 µl
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)
Processing	Manual (protocol for automated processing on QIAcube available)
Main sample type	FFPE tissue samples
Format	Spin column
Technology	Silica technology
Yield	Varies

Resources

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

Download Safety Data Sheets for QIAGEN product components.

Poster for download

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

Poster for download

FAQ

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

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

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

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

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

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

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