QIAseq FastSelect -5S/16S/23S Kit (24)

Bacterial rRNA removal during RNA-seq library preparation is significantly simplified by QIAGEN's QIAseq FastSelect –5S/16S/23S Kits, which use a novel method to remove highly abundant 5S, 16S and 23S rRNA. The kit was specifically designed for complex community microbial samples such as soil, water, stool and sludge, which contain numerous and diverse bacterial populations. Its rRNA coverage is based on SILVA 16S sequences (nearly 600,000 entries), SILVA 23S sequences (nearly 170,000 entries) and 5S rRNA database sequences (over 7200 entries), with in-silico modeling predicting >95% rRNA removal of all known 5S, 16S and 23S rRNA sequences.

The workflow removes rRNA in a single step without enzymatic digestion or hybrid-capture. After combining the QIAseq FastSelect reagent with total RNA (and fragmentation buffer, if desired), the reaction is stepwise cooled from 75°C to 25°C over 14 minutes, followed by a bead cleanup. The kit accommodates RNA inputs ranging from 20 ng to 1 µg and is compatible with low-quality, highly fragmented or high-quality full-length RNA, including FFPE and degraded samples.

The kit has been tested with QIAGEN, Illumina and NEB stranded RNA-seq library kits and integrates with all major methods of RNA-seq library preparation. Each lot is tested against predetermined specifications under QIAGEN's ISO-certified Quality Management System. In comparative testing on a gut microbiome sample, the kit removed over 90% of 5S/16S/23S rRNA and freed more than 85% of sequencing reads for non-rRNA gene expression analysis, increasing the number of detected genes (FPKM > 0) by approximately 50% compared with untreated controls.