Do not use for long read approaches.

This protocol is adapted from the protocol used in Adiconis et al., Nature Methods 2013 (http://doi.org/10.1038/nmeth.2483)

Goal : Method to ribodeplete samples from rRNAs using RNAseH nuclease.

Summary : DNA oligos are complementary to human rRNAs. They will anneal to the rRNA molecules. RNAseH will degrade RNA-DNA duplexes, leaving the RNA samples depleted from rRNAs

Tested on :Human, Mouse and Yeast (for 18s and 28s ribodepletion).

Cost : DNA oligonucleotides will be expensive the first time ordered, but will last for many rounds of ribodepletion. Once the DNA oligonucleotides have been purchased, the major cost comes from the RNAseH enzyme. In our hands, the cost is much lower than commercial kits, thus being scalable for high input samples where the ribodepleted material needs to be high.

Recommended for: NGS-based approaches. Do not use for Nanopore-based approaches as nuclease-based methods degrade RNA molecules.