Cat. No. / ID: 217504
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.
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 miRNeasy FFPE Kit allows purification of miRNA with total RNA for use in a variety of applications, such as quantitative, real-time RT-PCR.
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, CT 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, CT values were similar for both methods. CT values were similar or lower after purification using the miRNeasy FFPE Kit for all tissue types tested.
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 |