QIAseq UPXome RNA Lib Kit HMR (96)

This QIAGEN RNA library kit supports high-throughput transcriptome studies using an ultraplex workflow intended for human, mouse and rat samples. The kit features HMR-optimized depletion of abundant host rRNA and high-abundance transcripts while preserving coding and noncoding RNAs, and incorporates a streamlined, strand-specific protocol covering fragmentation, cDNA synthesis, end repair, adapter ligation and PCR. Integrated unique molecular identifiers (UMIs) support accurate duplicate handling and quantitation, and the workflow is Illumina compatible and suitable for automation or large cohort projects.

The QIAseq UPXome workflow supports a microplate-based approach, with cell lysis, rRNA depletion and reverse transcription performed in plate format, and a unique sample index incorporated into the cDNA from each sample so that samples can be pooled before amplification. Demonstrated use with Caco-2 and Hep G2 cell lines has shown that libraries can be prepared manually or semi-automated on the SPT Labtech mosquito HV genomics liquid handler, with sequencing performed on the Illumina NextSeq500 platform, quality assessment on an Agilent Tapestation D1000 screen tape and qPCR-based quantification. Data analysis has been reported using QIAGEN CLC Genomics Workbench and ExpressAnalyst. Reported mapping performance for libraries prepared at 0.5x reaction volumes with the semi-automated workflow ranged from 70 to 93% overall and 68 to 77% mapped to protein-coding regions for Caco-2 cells, and from 26 to 82% overall and 72 to 79% mapped to protein-coding regions for Hep G2 cells. Combining miniaturization with pre-amplification pooling yielded greater than 85% reduction in single-use plastic compared with a traditional RNA library preparation workflow.