RNA / miRNA
The PAXgene Tissue Systems consist of the PAXgene Tissue Fixation and Stabilization products for collection, transport, and storage of human tissue specimens and dedicated purification kits to isolate nucleic acids. The PAXgene Tissue RNA Kit provides optimized extraction of RNA from tissues fixed and stabilized in PAXgene Tissue Containers. The PAXgene Tissue miRNA Kit should be used to isolate total RNA (including miRNA). The RNA or miRNA can be either isolated from PAXgene Tissue fixed (PF) or PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue. The purification is performed manually in a spin-column format, though the PAXgene Tissue RNA Kit can also be automated on the QIAcube (see protocols).
Streamlined protocol for high yields of long RNA molecules
Together, the container and kit products provide a complete prenalytical solution for efficient purification of high-quality RNA and miRNA from PF or PFPE tissues for molecular analysis. These optimized solutions create appropriate conditions for RNA to bind to the silica membrane, allowing contaminants to be washed away. Treatment with DNase I removes any trace amounts of bound DNA, resulting in eluates of highly pure RNA.
Purification of low molecular weight RNAs
With the PAXgene Tissue RNA Kit procedure, size distribution of the purified RNA is comparable to that obtained by centrifugation on a CsCl gradient. RNAs < 200 nucleotides are selectively excluded. The modified chemistry of the PAXgene Tissue miRNA Kit was developed with optimized binding and washing conditions to ensure isolation of total RNA, including low molecular weight RNAs, such as 5.8S rRNA, 5S rRNA, tRNAs, and miRNAs.
High-quality RNAs for a range of downstream applications
Whether high or low molecular weight RNAs, the eluates resulting from the PAXgene Tissue RNA and miRNA Kits are of high quality and purity enabling their use in downstream applications including RT-PCR, quantitative, real-time RT-PCR, expression arrays and expression chip analysis, cDNA synthesis, RNA sequencing, RNase and S1 nuclease protection, Northern blotting, dot and slot blot analysis, and primer extension.