Qiagen

miRNeasy FFPE Kit (50)

miRNeasy FFPE Kit is intended for purification of microRNA and total RNA from formalin-fixed, paraffin-embedded tissue sections in molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

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

description

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. Procedures can be automated on the QIAcube Connect MDx.

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

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. Since RNA is eluted in a volume of just 14–30 µl, smaller reaction volumes are possible in downstream applications.

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.

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.

Study

Use of FFPE-derived DNA in next generation sequencing - NCBI

FEATURES & BENEFITS

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

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