miRNeasy FFPE Kit
For purification of microRNA and total RNA from formalin-fixed, paraffin-embedded tissue sections
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.
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 (see figures "Effective real-time RT-PCR quantification", "Efficient RNA purification from FFPE tissues" and Superior yields and performance).
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.
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 (see flowchart "miRNeasy FFPE procedure"). Since RNA is eluted in a volume of just 14–30 µl, smaller reaction volumes are possible in downstream 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.
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