Fast, simple mRNA library prep workflow for coding transcriptome analysis, combining cDNA synthesis and adapter tagging into a single step
Assay time
Hands-on time
Input quantity
Illumina Rapid mRNA Prep offers the fastest and easiest mRNA-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 gene expression data.
| Assay time | 3 hr |
|---|---|
| Automation capability | Liquid handling robot(s) |
| Description | A fast and easy workflow for coding transcriptome analysis |
| Hands-on time | < 1.5 hr |
| Input quantity | 1–1000 ng 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 | mRNA sequencing |
| Nucleic acid type | RNA |
| Specialized sample types | Not FFPE-compatible, Low-input samples |
| Species category | Mammalian, Mouse, Human, Rat |
| Strand specificity | Non-stranded |
| Technology | Sequencing |
| Variant class | Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants, Single nucleotide variants (SNVs) |
The complete Illumina Rapid mRNA Prep workflow requires Rapid RNA library prep reagents, mRNA capture kit, and indexes (each sold separately):
Illumina Rapid mRNA Prep enables preparation of sequencing-ready mRNA-Seq libraries for coding transcriptome analysis in 3 hr with less than 1.5 hr hands-on time.
| Instrument | Recommended number of samples | Read length |
|---|---|---|
| MiSeq i100 System, MiSeq i100 Plus System | 10–25M clusters per sample: 25M: 1–2, 50M: 1–5, 100M: 1–10 |
1 × 100 bp or 2 × 50 bp for gene expression, up to 2 × 100 bp for variant calling |
| NextSeq 1000 System, NextSeq 2000 System | 10M clusters per sample: P1: 10, P2: 40, P3: 120, P4: 180 25M clusters per sample: P1: 4, P2: 16, P3: 48, P4: 72 |
1 × 100 bp or 2 × 50 bp for gene expression, up to 2 × 100 bp for variant calling |
| NovaSeq X System, NovaSeq X Plus System | 10M clusters per sample: 1.5B: 150 (75/lane), 5B: 500 (62/lane), 10B: 1000 (125/lane), 25B: 2500 (312/lane) 25M clusters per sample: 1.5B: 60 (30/lane), 5B: 200 (25/lane), 10B: 400 (50/lane), 25B: 1000 (125/lane) |
1 × 100 bp or 2 × 50 bp for gene expression, up to 2 × 100 bp for variant calling |
mRNA sequencing (mRNA-Seq) using NGS can identify both known and novel transcripts, and measure transcript abundance.
Find out how to analyze RNA-Seq data with user-friendly software tools packaged in intuitive user interfaces designed for biologists.
Profile the transcriptome for a better understanding of biology.
| Illumina Rapid mRNA Prep | Illumina Stranded mRNA Prep | |
|---|---|---|
| Assay time | 3 hr | 6.5 hr |
| Automation capability | Liquid handling robot(s) | Liquid handling robot(s) |
| Description | A fast and easy workflow for coding transcriptome analysis | A simple, cost-effective solution for analysis of the coding transcriptome with precise strand information |
| Hands-on time | < 1.5 hr | < 3 hr |
| Input quantity | 1–1000 ng RNA | 25-1000 ng standard-quality total 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 | NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System |
| Method | mRNA sequencing | mRNA sequencing |
| Nucleic acid type | RNA | RNA |
| Specialized sample types | Not FFPE-compatible, Low-input samples | Not FFPE-compatible, Low-input samples |
| Species category | Mammalian, Mouse, Human, Rat | Mammalian, Bovine, Mouse, Human, Rat |
| 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 |
Library Prep and Array Kit Selector
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High library complexity with Illumina Rapid mRNA Prep
Illumina Rapid mRNA Prep data from 1, 10, 25, 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–32M clusters (2M–64M paired-end reads).
High mapping efficiency with Illumina Rapid mRNA Prep
Illumina Rapid mRNA Prep data shows high percentage of mapped reads across input amounts: 1, 10, 25, and 100 ng of universal human reference (UHR) RNA, downsampled to 20M clusters (40M paired-end reads).
Uniform coverage across transcripts with Illumina Rapid mRNA Prep
Illumina Rapid mRNA Prep data shows consistent normalized coverage distributed along the length of transcripts across RNA input amounts: 1, 10, 25, and 100 ng of universal human reference (UHR) RNA, downsampled to 20M clusters (40M paired-end reads).
High reproducibility across high and low RNA inputs
Illumina Rapid mRNA 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.
On-bead tagmentation library prep chemistry integrates DNA extraction, fragmentation, library prep, and library normalization.
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.
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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.
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Illumina mRNA Capture Kit (96 samples)
20158830
Includes reagents for capture and purification of polyadenylated mRNA for 96 samples. Compatible with Illumina Rapid RNA Prep, sold separately. for a complete Rapid mRNA library prep workflow.
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Illumina mRNA Capture Kit (16 samples)
20158829
Includes reagents for capture and purification of polyadenylated mRNA for 16 samples. Compatible with Illumina Rapid RNA Prep, sold separately. for a complete Rapid mRNA library prep workflow.
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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.
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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.
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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.
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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.
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Illumina Purification Beads 30 ml
20119944
Illumina Purification Beads 30 ml
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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.
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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.
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Step 1: Capture poly(A) RNA with the Illumina mRNA Capture 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. Illumina mRNA Capture is also available in 96-sample or 16-sample configurations.
Both methods start with oligo(dT) magnetic bead capture of poly(A) mRNA. Then Illumina Rapid mRNA Prep produces nonstranded libraries using tagmentation (3 hr total workflow time and < 1.5 hr hands-on time), while Illumina Stranded mRNA Prep produces stranded libraries using ligation (6.5 hr total workflow time and < 3 hr hands-on time).
Reagents for the MiSeq i100 Series feature easy-to-use cartridges and multiple flow cell configurations for flexible sequencing options.
Reagents for the NextSeq 1000/2000 System feature easy-to-use cartridges and multiple flow cell configurations for flexible sequencing options.
Reagent kits for the NovaSeq X Series include a reagent cartridge, buffer cartridge, flow cell, lyo insert, pre-load buffer, and library tube strip.
Fast, simple library prep workflow for RNA exome analysis, combining cDNA synthesis and adapter tagging into a single step.
A standalone kit allowing for ribosomal RNA and human globin mRNA removal in human, mouse, rat, and bacterial samples.
Fast, efficient depletion of undesirable host and pan-bacterial rRNA from complex microbial samples (eg stool) for metatranscriptomics research.
Fast, simple total RNA library prep workflow for whole-transcriptome analysis, combining cDNA synthesis and adapter tagging into a single step
A robust, highly scalable whole-transcriptome analysis (RNA-Seq) solution for a variety of species and sample types, including human, mouse, and formalin-fixed, paraffin-embedded (FFPE) tissue.
Rapid, simple RNA-Seq library prep workflow with custom enrichment, combining cDNA synthesis and adapter tagging into a single step.
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