Zymo-Seq RiboFree Total RNA-Seq Library Kit is the fastest, easiest RNA-Seq library prep kit available to make stranded, total RNA libraries. The automation-friendly protocol minimizes hands-on time to generate sequencer-ready, stranded, indexed libraries depleted of rRNA and globin in as little as 3.5 hours. Depletion of ribosomal RNAs (which can comprise as much as 90% of a total RNA sample) is completely integrated into the workflow, using a novel, probe-free technology that removes rRNA, globin, or any other over-abundant, repetitive transcript from any sample type or species. RiboFree Universal Depletion is validated across biological kingdom and phyla, including human, rodent, plant, and various prokaryotes, as well as RNA from a wide range of sample types including cells, tissue, FFPE tissues, and whole blood, eliminating the need to select sample-specific modules for different sample types. Just one total RNA-Seq library prep kit to deplete them all. Because the RiboFree Universal Depletion is probe-free, researchers using the Zymo-Seq RiboFree Total RNA Library Kit will avoid transcriptome profiling bias found in other methods. High concentrations of rRNA-binding oligonucleotides used in probe-based approaches also bind to a significant number of non-ribosomal targets. As much as 20% of the 20,000 protein-coding genes in the human genome are significantly biased by these methods. In contrast, the probe-free depletion developed by Zymo Research uses only abundant pre-existing transcripts for mismatch-free enzymatic removal of rRNA and globin, resulting in minimal off-target effects as low as 1.8% in all protein-coding transcripts. This total RNA-Seq library prep kit's protocol is fragmentation-free and works with up to 2 ug of undepleted total RNA, down to 50 ng of undepleted total RNA, or as little as 5 ng of ribosome-depleted RNA samples. Pre-mixed reagents are ready-to-use at every step, eliminating the need for manual master-mix preparation. Everything needed to make sequencing-ready libraries, including indexing primers, SPRI beads, and magnetic bead stand are included in the kit. It's RNA-Seq made simple.