Description
The RNeasy Pure mRNA Bead Kit allows highly specific polyA-oligo-dT–based purification of mRNA ready for next-generation sequencing (NGS) applications. By effectively depleting rRNA and non-adenylated, non-coding, as well as regulatory RNA from a wide variety of eukaryotic species, the kit enriches for mRNA, ensuring optimal use of expensive sequencing capacity and resources. With a preparation time of <45 min and convenient automation options, the RNeasy Pure mRNA Bead Kit offers unparalleled convenience by minimizing the hands-on time required to purify mRNA. Procedure can be automated on the QIAcube Connect. The RNeasy Pure mRNA Bead Kit formally known as the GeneRead Pure mRNA Kit contains identical chemistry.
Exceptional convenience and cost-effectiveness The RNeasy Pure mRNA Bead Kit contains spin columns and all necessary reagents and buffers for the isolation of pure poly A+ mRNA from total RNA preparations in <45 minutes. RNeasy Pure mRNA Bead Kit technology combines the convenient handling of magnetic particles with the specificity of oligo-dT hybridization. The streamlined protocol only requires short pretreatment steps. Products from other suppliers involve more time-consuming procedures as they generally require a second round of purification to achieve mRNA pure enough for NGS applications, thereby resulting in increased resources and costs. The RNeasy Pure mRNA Bead Kit is the only kit on the market that delivers mRNA of maximal purity with only one round of purification. Poly A+ mRNA purified with RNeasy Pure mRNA Bead Kit technology does not require further purification and is ready for downstream use in applications such as NGS. After purification with the RNeasy Pure mRNA Bead Kit, only meaningful reads are left — the majority of these reads are from polyA+ mRNA. Efficient removal of ribosomal RNA The RNeasy Pure mRNA Bead Kit is the only mRNA purification kit to remove rRNA, including mitochondrial rRNA, almost completely (Figures The RNeasy Pure mRNA Bead Kit outperforms mRNA purification kits from other suppliers and Specific and efficient mRNA enrichment and Table 1). Due to the kit’s stringent procedure, high mRNA purity is assured for increased reliability and reproducibility in sensitive downstream applications such as NGS. In addition, only small amounts of starting material, ranging from 5 μg down to 100 ng, are required. Maximal removal of rRNA Ribosomal RNA Removal (%) 28S 99.99 18S 99.99 5.8S 99.99 5S 99.99 16S mt 99.09 12S mt 99.78 Using the RNeasy Pure mRNA Bead Kit, 5 μg total RNA from Jurkat cells was purified. To determine the efficacy of removal of different types of rRNA, quantitative RT-PCR was performed.
RNeasy Pure mRNA Bead Kit technology combines the convenient handling of magnetic particles with the specificity of oligo-dT hybridization. Protocols require short pretreatment steps. Oligo-dT probes are covalently attached to the surface of Pure mRNA Beads. mRNA binds rapidly and efficiently to the oligo-dT probes on the Pure mRNA Beads in the presence of Buffer mRBB (see figure Convenient, streamlined procedure). mRNA bound to the magnetic particles is then efficiently washed. Two wash steps are used to considerably improve the purity of the mRNA. Highly pure mRNA is eluted in the elution buffer (Buffer OEB) provided with the kit. mRNA yields depend on sample input and sample storage.
The RNeasy Pure mRNA Bead Kit delivers mRNA of maximal purity with only one round of purification. The entire procedure takes <45 minutes (see figure Convenient, streamlined procedure). Minimal hands-on time required The majority of the RNeasy Pure mRNA Bead Kit procedure — from hybridization and capture through to RNA cleanup — can be fully automated on the QIAcube, affording maximal convenience and greater processing flexibility, and minimizing the hands-on time required.
