Fast, simple total RNA library prep workflow for whole-transcriptome analysis, combining cDNA synthesis and adapter tagging into a single step

Fast, simple total RNA library prep workflow for whole-transcriptome analysis, combining cDNA synthesis and adapter tagging into a single step

Illumina Rapid Total RNA Prep

Fast, simple total RNA library prep workflow for whole-transcriptome analysis, combining cDNA synthesis and adapter tagging into a single step

Illumina Rapid Total RNA Prep
Data sheet PDF 7 MB

3.5 hr

Assay time

< 1.5 hr

Hands-on time

1–1000 ng RNA or 10–1000 ng FFPE RNA

Input quantity

See full details in the specifications table

Learn
Order

Overview

Illumina Rapid Total RNA Prep offers the fastest and easiest total RNA-Seq library prep solution in the Illumina portfolio. A streamlined, 30-minute one-tube reaction combines cDNA synthesis and on-bead tagmentation. The rapid workflow supports low RNA sample inputs (1–1000 ng) and delivers high-accuracy nonstranded whole-transcriptome data.

Learn about on-bead tagmentation

Key features

Fast, simplified library prep

The entire simplified workflow, including rRNA depletion and PCR steps, requires only 3.5 hours to generate a sequencing-ready total RNA library.

Optimized performance

Removes prokaryotic and eukaryotic ribosomal RNA (rRNA) and globin RNA from multiple species in a single reaction to allow rich transcriptome analysis.

High-quality data from low input samples

Reproducible results from as little as 1 ng of high-quality RNA or 10 ng FFPE* RNA while achieving uniform coverage.

*FFPE, formalin-fixed, paraffin-embedded samples.

Specifications

Assay time	3.5 hr
Automation capability	Liquid handling robot(s)
Description	A fast and easy workflow for whole-transcriptome analysis.
Hands-on time	< 1.5 hr
Input quantity	1–1000 ng RNA or 10–1000 ng FFPE RNA
Instruments	NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NextSeq 550Dx in Research Mode, NovaSeq X System, NovaSeq 6000Dx in Research Mode, NovaSeq 6000 System, NovaSeq X Plus System, MiSeq i100 System, MiSeq i100 Plus System
Method	Whole-transcriptome sequencing, Metatranscriptome sequencing
Nucleic acid type	RNA
Specialized sample types	Microbiome samples, Low-input samples, FFPE tissue
Species category	Mouse, Human, Rat, Bacteria
Species details	Best for organisms with well-annotated genomes; Ribo-Zero Plus depletes abundant transcripts from multiple species including: human cytoplasmic and mitochondrial rRNA, mouse rRNA, rat rRNA, bacteria ( E. coli and B. subtilis) rRNA, and human beta globin transcripts; Ribo-Zero Plus Microbiome depletes abundant transcripts from multiple species including: those depleted in Ribo-Zero Plus as well as diverse microbial species and ATCC MSA-2002, MSA-2005, and MSA-2006. May support other species with custom rRNA probe design spike-in to RiboZero Plus.
Strand specificity	Non-stranded
Technology	Sequencing
Variant class	Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants, Single nucleotide variants (SNVs)

Required Products

The complete Illumina Rapid Total RNA Prep workflow requires Rapid RNA library prep reagents, Ribo-Zero Plus rRNA depletion kit, and indexes (each sold separately):

Illumina Rapid RNA Prep: contains library prep reagents (96 sample or 16 sample kits available)
Ribo-Zero Plus rRNA Depletion Kit OR Ribo-Zero Plus Microbiome Depletion Kit: contains reagents for removal of abundant RNAs (96 sample or 16 sample kits available)
Illumina DNA/RNA UD Index Set, Tagmentation: each set of 96 indexes supports index labeling of 96 samples

/ Results

Applications

Illumina Rapid Total RNA Prep enables preparation of sequencing-ready total RNA-Seq libraries for whole-transcriptome analysis in 3.5 hr with less than 1.5 hr hands-on time.

Example workflow

Prep

Illumina Rapid Total RNA Prep

Sequence

Analyze

Project recommendations

Instrument	Recommended number of samples	Read length
MiSeq i100 Plus System	Samples per run (50M clusters per sample): 50M: 1, 100M: 1–2 (25M clusters/sample) 50M: 2, 100M: 4	1 × 100 bp, or 2 × 50 bp for use with 100 cycle kits
NextSeq 2000 System	Samples per run (50M clusters per sample): P1: 2, P2: 8, P3: 24, P4: 36 (25M clusters/sample) P1: 4, P2: 16, P3: 48, P4: 72	1 × 100 bp, or 2 × 50 bp for use with 100 cycle kits
NovaSeq X System, NovaSeq X Plus System	Samples per flow cell (50M clusters per sample): 1.5B: ~30, 5B: ~100, 10B: ~200, 25B: ~520	1 × 100 bp, or 2 × 50 bp for use with 100 cycle kits

Instrument

Recommended number of samples

Read length

MiSeq i100 Plus System

Samples per run (50M clusters per sample): 50M: 1, 100M: 1–2

(25M clusters/sample) 50M: 2, 100M: 4

1 × 100 bp, or 2 × 50 bp for use with 100 cycle kits

NextSeq 2000 System

Samples per run (50M clusters per sample): P1: 2, P2: 8, P3: 24, P4: 36

(25M clusters/sample) P1: 4, P2: 16, P3: 48, P4: 72

1 × 100 bp, or 2 × 50 bp for use with 100 cycle kits

NovaSeq X System, NovaSeq X Plus System

Samples per flow cell (50M clusters per sample): 1.5B: ~30, 5B: ~100, 10B: ~200, 25B: ~520

1 × 100 bp, or 2 × 50 bp for use with 100 cycle kits

Related applications and methods

Total RNA Sequencing

Analyze both coding RNA and multiple forms of noncoding RNA for a comprehensive view of the transcriptome.

RNA Sequencing Data Analysis

Find out how to analyze RNA-Seq data with user-friendly software tools packaged in intuitive user interfaces designed for biologists.

Transcriptomics

Profile the transcriptome for a better understanding of biology.

Documentation

Product literature

Illumina Rapid Total RNA Prep
Data sheet PDF 7 MB

Support documentation

Illumina Rapid Total RNA Prep with RiboZero Plus Product Documentation

Illumina Rapid Total RNA Prep with RiboZero Plus Documentation

All Illumina Rapid Total RNA Prep with RiboZero Plus Support

Compare

	Illumina Rapid Total RNA Prep	Illumina Stranded Total RNA Prep with Ribo-Zero Plus or Ribo-Zero Plus Microbiome
Assay time	3.5 hr	~7 hr
Automation capability	Liquid handling robot(s)	Liquid handling robot(s)
Description	A fast and easy workflow for whole-transcriptome analysis.	For the preparation of whole-transcriptome sequencing-ready libraries from low RNA inputs and low-quality samples that supports a wide range of RNA-Seq applications. Includes Ribo-Zero Plus for single-tube depletion of rRNA and globin RNA from multiple species to facilitate rich transcriptome analyses.
Hands-on time	< 1.5 hr	< 3 hr
Input quantity	1–1000 ng RNA or 10–1000 ng FFPE RNA	1–1000 ng total RNA for standard-quality RNA samples. 10 ng total RNA minimum recommended for optimal performance and FFPE samples
Instruments	NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NextSeq 550Dx in Research Mode, NovaSeq X System, NovaSeq 6000Dx in Research Mode, NovaSeq 6000 System, NovaSeq X Plus System, MiSeq i100 System, MiSeq i100 Plus System	NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System
Method	Whole-transcriptome sequencing, Metatranscriptome sequencing	Whole-transcriptome sequencing, Metatranscriptome sequencing
Nucleic acid type	RNA	RNA
Specialized sample types	Microbiome samples, Low-input samples, FFPE tissue	Microbiome samples, Low-input samples, FFPE tissue
Species category	Mouse, Human, Rat, Bacteria	Mouse, Human, Rat, Bacteria
Species details	Best for organisms with well-annotated genomes; Ribo-Zero Plus depletes abundant transcripts from multiple species including: human cytoplasmic and mitochondrial rRNA, mouse rRNA, rat rRNA, bacteria ( E. coli and B. subtilis) rRNA, and human beta globin transcripts; Ribo-Zero Plus Microbiome depletes abundant transcripts from multiple species including: those depleted in Ribo-Zero Plus as well as diverse microbial species and ATCC MSA-2002, MSA-2005, and MSA-2006. May support other species with custom rRNA probe design spike-in to RiboZero Plus.	Ribo-Zero Plus depletes abundant transcripts from multiple species including: human cytoplasmic and mitochondrial rRNA, mouse rRNA, rat rRNA, bacteria ( E. coli and B. subtilis) rRNA, and human beta globin transcripts Ribo-Zero Plus Microbiome depletes abundant transcripts from multiple species including: those depleted in Ribo-Zero Plus as well as diverse microbial species and ATCC MSA-2002, MSA-2005, and MSA-2006
Strand specificity	Non-stranded	Stranded
Technology	Sequencing	Sequencing
Variant class	Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants, Single nucleotide variants (SNVs)	Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants

Selection tools:

Library Prep and Array Kit Selector

Find the right sequencing library preparation kit or microarray for your needs. Filter by method, species, and more. Compare, share, and order kits.

Figures

Effective depletion of rRNA using Ribo-Zero Plus

Percent of total reads that align to abundant RNA in depleted (Ribo-Zero Plus rRNA Depletion Kit) and undepleted samples: 10 ng blood RNA, 10 ng universal human reference (UHR) RNA, and 10 ng universal human brain reference (UHBR) RNA.

Effective depletion of globin RNA using Ribo-Zero Plus

Normalized transcripts per million (TPM) for globin genes: HBA1/HBA2 (alpha), HBB (beta), and HBG1/HBG2 (gamma) for 10 ng depleted (Ribo-Zero Plus rRNA Depletion Kit) and undepleted universal human reference (UHR) RNA. HBG1/HBG2 are fetal-specific genes, not expected to be expressed in UHR samples.

High library complexity with Illumina Rapid Total RNA Prep

Illumina Rapid Total RNA Prep data from 1 ng, 10 ng, and 100 ng of universal human reference (UHR) RNA shows the number of unique genes detected (≥ 5 mapped reads) across sequence depths downsampled to 1M–50M clusters (2M–100M paired-end reads).

High mapping efficiency with Illumina Rapid Total RNA Prep

Illumina Rapid Total RNA Prep data shows high percentage of mapped reads across input amounts: 1 ng, 10 ng, and 100 ng of universal human brain reference (UHBR) RNA and universal human reference (UHR) RNA.

Uniform coverage across transcripts with Illumina Rapid Total RNA Prep

Illumina Rapid Total RNA Prep data shows consistent normalized coverage distributed along the length of transcripts across RNA input amounts: 1 ng, 10 ng, and 100 ng of universal human brain reference (UHBR) RNA and universal human reference (UHR) RNA.

High reproducibility across high and low RNA inputs

Illumina Rapid Total RNA Prep shows high correlation of normalized gene counts between 1 ng and 100 ng input amounts (R² = 0.99). Transcripts per million (TPM) generated by DRAGEN RNA v4.4.

Resources

On-bead tagmentation

Library prep (5)

Illumina Rapid RNA Prep, (S) Tagmentation (96 samples)

20158824

Includes reagents for preparing 96 samples of Rapid RNA libraries with Illumina Purification Beads. UD Indexes sold separately. Compatible with mRNA Capture kit for complete mRNA library prep workflow or RiboZero Plus kits for complete Total RNA library prep workflow or RNA Fast Hyb Enrichment kits for complete RNA Enrichment workflow, each sold separately.

List Price:

Discounts:

Illumina Rapid RNA Prep, (S) Tagmentation (16 samples)

20158823

Includes reagents for preparing 16 samples of Rapid RNA libraries with Illumina Purification Beads. UD Indexes sold separately. Compatible with mRNA Capture kit for Rapid mRNA library prep workflow or RiboZero Plus kits for Rapid Total RNA library prep workflow or Fast Hyb Enrichment kits for Rapid RNA Enrichment library prep workflow, each sold separately.

List Price:

Discounts:

Illumina Ribo-Zero plus rRNA Depletion Kit (96 Samples)

20037135

Includes Ribo-Zero Plus depletion module for depletion of ribosomal RNA from human, mouse, rat, and bacteria and globin transcripts for 96 samples. Purchase purification beads, library prep reagents and index adapters separately. For complete kit, including library prep reagents, order the Illumina Stranded Total RNA with Ribo-Zero Plus kit.

List Price:

Discounts:

Illumina® Ribo-Zero Plus rRNA Depletion Kit (16 Samples)

20040526

List Price:

Discounts:

Illumina® Ribo-Zero Plus rRNA Microbiome depletion kit (96 Samples)

20072062

Includes Ribo-Zero Plus Microbiome depletion module for depletion of ribosomal RNA from diverse bacteria commonly found in microbiomes and targets included in the Ribo-Zero Plus kit for 96 samples. For complete kit including library prep reagents, order the Illumina Stranded Total RNA with Ribo-Zero Plus Microbiome kit.

List Price:

Discounts:

Index adapters (4)

Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples)

20091654

Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.

List Price:

Discounts:

Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples)

20091656

Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.

List Price:

Discounts:

Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples)

20091658

Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.

List Price:

Discounts:

Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples)

20091660

Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.

List Price:

Discounts:

Accessory products (3)

Illumina Purification Beads 30 ml

20119944

Illumina Purification Beads 30 ml

List Price:

Discounts:

Illumina Purification Bead, 100mL

20060057

Includes one, 100-ml container of beads for library clean-up and size selection. Purchase library prep and enrichment reagents, probe panels, and indexes separately.

List Price:

Discounts:

Illumina Purification Bead, 400mL

20060058

Includes four, 100-ml container of beads for library clean-up and size selection. Purchase library prep and enrichment reagents, probe panels, and indexes separately.

List Price:

Discounts:

Showing of

Selection summary

Product

Qty

Unit price

Total ( items)

You have not selected any items yet.

Please add these products to your cart before leaving the page to avoid losing them.

FAQs

Step 1: Remove abundant RNA species with the Ribo-Zero Plus rRNA Depletion Kit

Step 2: Illumina Rapid RNA Prep combines cDNA synthesis and on-bead tagmentation in a single-tube, 30-min reaction

Step 3: PCR adds index adapters

Step 4: Clean up with included Illumina Purification Beads

Illumina Rapid RNA Prep is available in 96-sample and 16-sample configurations. Ribo-Zero Plus rRNA Depletion Kits are also available in 96-sample or 16-sample configurations.

Both methods start with Ribo-Zero Plus depletion of abundant RNA species. Then Illumina Rapid Total RNA Prep produces nonstranded libraries using tagmentation (3.5 hr total workflow time and < 1.5 hr hands-on time), while Illumina Stranded Total RNA Prep produces stranded libraries using ligation (7 hr total workflow time and < 3 hr hands-on time).

/ Results

Frequently purchased
Other products to consider

Speak with a specialist

Reach out for information about our products and services, or get answers to questions about our technology.