Ribo-Zero® rRNA Removal Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
This kit support for intact and partially degraded small samples, requiring input of ≥ 1 µg total RNA. Kit supports rRNA depletion from 1–5 µg total RNA samples		Downstream sequencing data that contains complete transcriptome of coding and noncoding RNA species.

Upstream tips
This kit support for intact and partially degraded small samples, requiring input of ≥ 1 µg total RNA. Kit supports rRNA depletion from 1–5 µg total RNA samples
Downstream tips
Downstream sequencing data that contains complete transcriptome of coding and noncoding RNA species.

Publication protocol

RNA was extracted from frozen brain tissues using Tissue Lyser LT and RNeasy Mini Kit (Qiagen, Hilden, Germany) following the manufacturer’s instruction. RIN (RNA integrity) and DV200 were measured with RNA 6000 Pico Assay using Bioanalyzer 2100 (Agilent Technologies). The RIN is determined by the software on the Bioanalyzer taking into account the entire electrophoretic trace of the RNA including the presence or absence of degradation products. The DV200 value is defined as the percentage of nucleotides > 200 nt. RIN and DV200 for all the samples can be found on Additional file 1: Table S1. The yield of each sample is determined by the Quant-iT RNA Assay (Life Technologies) on the Qubit Fluorometer (Fisher Scientific). The complementary DNA (cDNA) library was prepared with the TruSeq Stranded Total RNA Sample Prep with Ribo-Zero Gold kit (Illumina) and then sequenced by HiSeq 4000 (Illumina) using 2 × 150 paired-end reads at McDonnell Genome Institute, Washington University in St. Louis with a mean of 58.14 ± 8.62 million reads. Number of reads and other quality control (QC) metrics can be found in Additional file 1: Table S1.

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Manufacturer protocol

Download the product protocol from Illumina for Ribo-Zero® rRNA Removal Kit below.

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