AllPrep DNA/mRNA Nano (12)

From QIAGEN, this magnetic-bead-based kit enables the simultaneous purification of genomic DNA (gDNA) and messenger RNA (mRNA) from low-biomass samples. A two-step magnetic-bead workflow maximises mRNA capture while isolating high-molecular-weight DNA for multiomics studies.

The system is designed for low-biomass inputs such as circulating tumour cells (CTCs), rare cells, fetal cells, stem cells and T-cells, with sample amounts down to 1 cell. mRNA is captured using oligo-dT magnetic beads and gDNA using DNA-affinity beads, with manual processing on a magnetic rack such as the AdnaMag-S. The procedure avoids phenol, chloroform and alcohol precipitation. Genomic DNA obtained with the kit has an average length of 60–100 kb depending on homogenisation conditions and a high molecular weight of up to 100 kb.

Downstream uses include PCR, multiplex PCR, dPCR, pyrosequencing, NGS, RT-PCR, real-time RT-PCR, cDNA synthesis, RNA-seq, and transcriptome and genome profiling, as well as the AdnaTest workflow for molecular characterisation of CTCs. The elution volume is 25 µl. The kit allows the parallel processing of multiple samples in less than 75 minutes, with a time per prep of 75 min. In a spiking experiment with six samples in AllPrep DNA/mRNA Nano lysis buffer, each spiked with 5 MCF-7 cells and 5 LNCaP95 cells, a range of 7.9–11.9 ng/µl (median 10 ng/µl) of mRNA was isolated and gDNA was detected within 28.3–29.6 cycles (median Cq 28.8).