Illumina Stranded Total RNA Library Prep Training
Illumina Total RNA Prep with Ribo-Zero Plus provides exceptional performance for the analysis of coding and multiple forms of noncoding RNA. Gain high-confidence discovery of alternative transcripts, gene fusions, and allele-specific expression.
Illumina Stranded Total RNA Prep with Ribo-Zero Plus Microbiome effectively depletes unwanted host and bacterial rRNA from thousands of bacterial strains commonly found in the gut for high-quality metatranscriptomics applications.
Read the data sheet to learn more about metatranscriptomics applications.
Illumina Total RNA Prep with Ribo-Zero Plus is based on TruSeq ligation technology, which has been cited in over 9,926 publications since 2011. This ligation method results in high coverage uniformity, precise strand information, and robust and reliable library preparation even from degraded samples.
Learn more about ligation technology
Illumina Total RNA Prep with Ribo-Zero Plus supports a broad range of RNA inputs, from 1 ng to 1000 ng1. It’s compatible with various sample types, including formalin-fixed paraffin-embedded (FFPE) and other low-quality samples. The included Ribo-Zero Plus removes abundant RNA from multiple species, including human, mouse, rat, bacteria, and epidemiology samples.
Illumina Stranded Total RNA Prep with Ribo-Zero Plus Microbiome provides robust depletion of abundant rRNA in complex microbial samples, including stool samples, for metatranscriptomic studies. This kit accommodates a broad range of RNA inputs from 25 ng to 1000 ng2, and depletes rRNA from human, mouse, rat, and thousands of bacterial species.
Prepare libraries for Illumina sequencing in less than a day. Illumina Qualified Methods are available on a range of automation platforms through our partners.
Explore library prep automation solutions
Multiplex up to 384 samples and sequence in a single run. The number of samples sequenced per run depends on the throughput capacity of the sequencer.
1This input range is applicable to high-quality RNA. We recommend 10 ng input for best results and for FFPE or degraded samples.
2Minimum recommended input for Ribo-Zero Plus Microbiome is 25 ng.
|Illumina Stranded Total RNA Prep with Ribo-Zero Plus or Ribo-Zero Plus Microbiome||TruSeq Stranded Total RNA||TruSeq Stranded Total RNA with Ribo-Zero Globin|
|Assay Time||~7 hours||11.5 hours||11.5 hours|
|Automation Capability||Liquid Handling Robots||Liquid Handling Robots||Liquid Handling Robots|
|Content Specifications||Captures coding RNA plus multiple forms of non-coding RNA||Captures coding RNA plus multiple forms of non-coding RNA||Captures coding RNA plus multiple forms of non-coding RNA|
|Description||For whole-transcriptome sequencing studies of multiple sample types. Includes Ribo-Zero plus module for single-tube depletion of abundant transcripts from multiple species||For whole-transcriptome sequencing studies. The Total RNA version depletes cytoplasmic rRNA while the Total RNA Gold version depletes cytoplasmic and mitochondrial rRNA.||For whole-transcriptome sequencing studies of blood-derived RNA. Depletes cytoplasmic and mitochondrial rRNA plus globin mRNA.|
|Hands-On Time||< 3 hours||5.5 hours||5.5 hours|
|Input Quantity||1-1000 ng total RNA for standard-quality RNA samples. 10ng total RNA minimum recommended for optimal performance and FFPE samples||0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization.||0.1 – 1 ug high-quality purified total RNA from blood|
|Mechanism of Action||Enzymatic rRNA depletion, ligation-based addition of adapters and indexes||Bead-based rRNA depletion, cDNA synthesis, and PCR||Bead-based rRNA depletion, cDNA synthesis, and PCR|
|Method||Metatranscriptome Sequencing, Whole-Transcriptome Sequencing||Whole-Transcriptome Sequencing||Whole-Transcriptome Sequencing|
|Multiplexing||Up to 384 Unique Dual Indexes (UDIs)||Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together, High-throughput kits: Prepare 96 indexes samples using either combinatorial dual indexes or unique dual indexes (available soon)||High-throughput kits: Prepare 96 indexes samples using either combinatorial dual indexes or unique dual indexes (available soon), Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together|
|Species Category||Bacteria, Human, Mouse, Rat||Human, Mouse, Rat||Human, Mouse, Rat|
Data Sheet | PDF | 6 versions
Brochure | PDF 1 MB
Illumina library preparation solutions
Brochure | PDF 2 MB
NextSeq 1000 and NextSeq 2000 RNA sequencing solution
Application Note | PDF < 1 MB
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Best practices for read trimming for Illumina Stranded mRNA and Total RNA workflows
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Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Consumables & Equipment Documentation
Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Checklist Documentation
Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Reference Guide Documentation
Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Documentation
Custom Protocol Selector
Generates customized, end-to-end instructions
All Illumina Stranded Total RNA Prep Ligation with Ribo-Zero Plus Support
See how Drs. Tonya Ward, Emily Hollister, and Le François of Diversigen and DNA Genotek developed workflows for metatranscriptomic studies and read their views for this research area’s potentially impactful future.Read Customer Spotlight
In this webinar, you'll hear from three leading experts in the field as they share their insights and key considerations on sample collection and processing, rRNA depletion, and bioinformatic analysis.Watch Webinar