Description
The QIAseq Universal Metagenome Library Kit converts all nucleic acid types into a single sequencing-ready library in a unified process. Starting from RNA, DNA or total nucleic acid, the QIAseq Universal Metagenome Kit will generate libraries from all microbial targets – viral, bacterial or fungal. To improve sensitivity, the workflow has built in mammalian host rRNA and globin RNA depletion during reverse transcription using QIAseq FastSelect technology. The workflow scales from up to 384 samples and is compatible with all Illumina platforms. For analysis, libraries integrate with CLC Genomics Workbench to identify organisms, variants, phylogeny and integration sites. Each lot is tested under ISO-certified quality programs.
The QIAseq Universal Metagenome Library Kit delivers consistent library quality across complex sample types. Libraries show a tight fragment size distribution, typically 250-350 bp, with minimal adapter dimers after cleanup. This size range supports stable cluster generation and even read coverage across genomes. Host rRNA and globin RNA depletion occurs during reverse transcription. This step increases usable microbial reads in samples with high host background, such as human and environmental matrices. Depending on sample type, QIAseq Bacterial FastSelect or other FastSelect targets can be used. Libraries perform consistently on all major Illumina platforms using paired-end sequencing. The recommended setup uses 149 bp paired-end reads and dual indexing for accurate demultiplexing. As a starting point, allocating 10–20 million read clusters per sample supports broad metagenome detection. Quantification by qPCR confirms accurate library concentration and balanced pooling. When paired with proper quantification, the kit delivers reproducible cluster density and high percentages of reads passing filter.
The QIAseq Universal Metagenome Library Kit uses a unified chemistry to convert mixed microbial nucleic acids into sequencing-ready libraries. First, total RNA in the sample is reverse transcribed using random priming. During this step, mammalian host rRNA and globin RNA are depleted. The generated single stranded cDNA then undergoes second strand synthesis to create double stranded cDNA. This part of the process will also convert ssDNA into double stranded cDNA. This double stranded cDNA, together with the DNA present from the sample, serves as input for library construction. After bead cleanup, all nucleic acids enter the same downstream process. Next, the QIAseq FX DNA Library Prep Kit chemistry performs enzymatic fragmentation, end repair, and A-addition in one tube. Illumina-compatible adapters with Unique Dual Indexes are ligated to both ends of each fragment. Following library amplification and cleanup, the result is a high-quality, indexed library. The final libraries are compatible with all Illumina sequencers and downstream metagenomic analysis. This procedure delivers sequencing-ready metagenome libraries from diverse sample types.
