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RNA Sequencing Sample Preparation |
Next-generation sequencing has transformed gene expression profiling and other RNA expression analysis studies. With an increasing number of researchers employing next-generation sequencing, and with the evolution of sequencing platforms, it is essential to use robust and streamlined methods for sample preparation. The best next-generation sequencing data begin with optimal samples.
RNA Sequencing Overview
Next-generation sequencing studies require robust and streamlined methods for sample preparation. The best next-generation sequencing data begin with optimal samples. Click the tabs above to learn more and see all available products.
RNA Isolation
Although many methods are available for purifying RNA from any particular source, obtaining RNA of the highest purity is the best assurance that the downstream manipulations will be successful. Choose from various formats, including PureLink, mirVana, RecoverAll, and MagMAX kits to isolate total RNA, then enrich for the subpopulation of interest.
| Order now | Order now | Order now | Order now | Order now | |
| Simple, reliable, rapid method | 30 min isolation from most samples | HTP format, ideal for stringent applications | Best for next-generation sequencing of mRNA | High-throughput purification of FFPE samples | |
|---|---|---|---|---|---|
| PureLink RNA Mini Kit | mirVana miRNA Isolation Kit (with or without phenol) | MagMAX for Microarrays Total RNA Isolation Kit | Dynabeads mRNA DIRECT Micro Kit | MagMAX FFPE Total Nucleic Acid Isolation Kit | |
| RNA types isolated for sequencing | Large RNA molecules only (mRNA and rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) | mRNA only | total RNA, microRNA, gDNA |
| Isolation method | Fast, convenient silica column | Highest purity and convenience; includes organic extraction and silica column | Scalable, flexible format for highest purity; includes organic extraction and magnetic beads | Fast magnetic bead capture directly from microsized samples | Xylene-free, scalable, flexible format with magnetic beads |
| Prep time | 20 min | 30 min | <1 hr | 15 min | 3 hr (for 96 preps) |
| Amount of starting material | 10 mg to 100 mg of tissue Up to 5 x 107 cells | Up to 100 mg of tissue Up to 1 x 107 cells | Up to 100 mg of tissue Up to 1 x 107 cells | Up to 500 ng purified Total RNA | 2 x 10 micron FFPE sections |
| Prep size | 50 preps | 40 preps | 96 preps | 2 mL oligo dT beads (sufficient for 100 mRNA preps) | 96 preps |
RNA Enrichment
To maximize the effectiveness of sequencing reagents, it is often necessary to enrich the sample for specific regions or targets of interest. This enrichment helps eliminate nonspecific artifacts, making it possible to better analyze smaller changes in genomes or transcriptomes and less abundant sequences. Ambion RiboMinus technology is designed to enrich the whole spectrum of RNA transcripts, regardless of their polyadenylation status or the presence of a 5' cap, by selectively depleting rRNA. The RiboMinus method removes the vast majority of rRNA (up to 99.9%) to allow greater interrogation of the less abundant transcripts.
Learn more about RiboMinus technology
| Order now | Order now | |
| A novel and efficient method utilizing Locked Nucleic Acids (LNA) to remove up to 99% rRNA | A novel and efficient method utilizing Locked Nucleic Acids (LNA) to remove up to 99% rRNA | |
|---|---|---|
| RiboMinus Eukaryote Kit for RNA-Seq | RiboMinus Plant Kit for RNA-Seq | |
| Platform | Diagnostic | Diagnostic |
| Starting material | 1–10 µg total RNA 100–500 ng total RNA (low-input protocol) | 1–10 µg total RNA |
| RiboMinus RNA yield | 1–2 µg from 10 µg human total RNA ~100 ng from 500 ng human total RNA (low-input protocol) | 1–2 µg from 10 µg human total RNA |
| Probe specificity | Eukaryotic | Plant |
| Cat. No. | A1083708 - 8 preps | A1083808 - 8 preps |
RNA Library Construction
Library construction, template preparation, and sequencing are at the heart of the sequencing workflow. Utilization of sequencing controls and methods for data analysis complete the workflow and provide useful annotated data. Generate cDNA by reverse transcription from adaptors ligated to the ends of RNA, then amplify the cDNA using primers complementary to the adaptors, and purify it.
| Order now | Order now | Order now | |
| Prepare representative cDNA libraries for strand-specific RNA sequencing | A Great Solution for NGS | The Ultimate Solution for NGS | |
|---|---|---|---|
| Ion Total RNA-Seq Kit v2 | SOLiD Total RNA-Seq Kit | Ambion RNA-Seq Library Construction Kit | |
| Platform | Ion Torrent PGM | SOLiD 5500 Analyzer | Illumina |
| RNA input type | rRNA-depleted total RNA, poly(A) RNA, or miRNA | Total RNA, rRNA-depleted total RNA, poly(A) RNA, or miRNA | Total RNA, rRNA-depleted total RNA, poly(A) RNA, or miRNA |
| RNA input range | Small RNA: total RNA containing 1–100 ng of miRNA Whole transcriptome: 100 ng total RNA (~1 ng poly(A) or ~25 ng rRNA-RNA | Small RNA: total RNA containing 1–100 ng of miRNA Whole transcriptome: 100 ng poly(A), 200 ng | Small RNA: total RNA containing 1–100 ng of miRNA Whole transcriptome: 100 ng poly(A) or 200 ng rRNA-depleted total RNA, or 200 ng total RNA |
| Barcodes available | Yes | 96 | No |
| Cat. No. | 4475936 - 12 reaction kit | 4445374 - 12 reaction kit | 4454073 - 12 reaction kit |
RNA Library Controls
Library construction, template preparation, and sequencing are at the heart of the sequencing workflow. Utilization of sequencing controls and methods for data analysis complete the workflow and provide useful annotated data.
Learn more about ERCC Controls
| Order now | Order now | |
| 2 tubes containing preformulated mixes of the same 92 RNA transcripts but in different ratios, for measuring dynamic range, assessing bias, and confirming expression fold changes | Single tube containing a preformulated mix of 92 RNA transcripts for measuring dynamic range and assessing bias | |
|---|---|---|
| ERCC ExFold RNA Spike-In Mixes (2 x 10 µL) | ERCC RNA Spike-In Mix (1 x 10 µL) | |
| Top Seller | ||
| Compatible sample types | Eukaryotic total RNA, rRNA-depleted total RNA, or poly(A) RNA | Eukaryotic total RNA, rRNA-depleted total RNA, or poly(A) RNA |
| Dynamic range | 6 logs | 6 logs |
| Volume | 10 µL (2 tubes) | 10 µL |
| Number of reactions | Spike-in controls diluted based on sample type and concentration | Spike-in controls diluted based on sample type and concentration |
RNA Barcoding
Sets of 16 barcodes are available for use with the SOLiD Total RNA-Seq Kit or the SOLiD SAGE Kit with Barcoding Adaptor Module. Barcodes cost-effectively increase the number of samples that can be analyzed in a single sequencing run. Barcodes assign a unique identifier to templated beads made from a single library. Once assigned, the RNA libraries can be pooled and sequenced.
Cat. No. | Product |
RNA Sequencing Overview
Next-generation sequencing studies require robust and streamlined methods for sample preparation. The best next-generation sequencing data begin with optimal samples. Click the tabs above to learn more and see all available products.
RNA Isolation
Although many methods are available for purifying RNA from any particular source, obtaining RNA of the highest purity is the best assurance that the downstream manipulations will be successful. Choose from various formats, including PureLink, mirVana, RecoverAll, and MagMAX kits to isolate total RNA, then enrich for the subpopulation of interest.
| Order now | Order now | Order now | Order now | Order now | |
| Simple, reliable, rapid method | 30 min isolation from most samples | HTP format, ideal for stringent applications | Best for next-generation sequencing of mRNA | High-throughput purification of FFPE samples | |
|---|---|---|---|---|---|
| PureLink RNA Mini Kit | mirVana miRNA Isolation Kit (with or without phenol) | MagMAX for Microarrays Total RNA Isolation Kit | Dynabeads mRNA DIRECT Micro Kit | MagMAX FFPE Total Nucleic Acid Isolation Kit | |
| RNA types isolated for sequencing | Large RNA molecules only (mRNA and rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) | Small & large RNA molecules (microRNA, tRNA, mRNA, rRNA) | mRNA only | total RNA, microRNA, gDNA |
| Isolation method | Fast, convenient silica column | Highest purity and convenience; includes organic extraction and silica column | Scalable, flexible format for highest purity; includes organic extraction and magnetic beads | Fast magnetic bead capture directly from microsized samples | Xylene-free, scalable, flexible format with magnetic beads |
| Prep time | 20 min | 30 min | <1 hr | 15 min | 3 hr (for 96 preps) |
| Amount of starting material | 10 mg to 100 mg of tissue Up to 5 x 107 cells | Up to 100 mg of tissue Up to 1 x 107 cells | Up to 100 mg of tissue Up to 1 x 107 cells | Up to 500 ng purified Total RNA | 2 x 10 micron FFPE sections |
| Prep size | 50 preps | 40 preps | 96 preps | 2 mL oligo dT beads (sufficient for 100 mRNA preps) | 96 preps |
RNA Enrichment
To maximize the effectiveness of sequencing reagents, it is often necessary to enrich the sample for specific regions or targets of interest. This enrichment helps eliminate nonspecific artifacts, making it possible to better analyze smaller changes in genomes or transcriptomes and less abundant sequences. Ambion RiboMinus technology is designed to enrich the whole spectrum of RNA transcripts, regardless of their polyadenylation status or the presence of a 5' cap, by selectively depleting rRNA. The RiboMinus method removes the vast majority of rRNA (up to 99.9%) to allow greater interrogation of the less abundant transcripts.
Learn more about RiboMinus technology
| Order now | Order now | |
| A novel and efficient method utilizing Locked Nucleic Acids (LNA) to remove up to 99% rRNA | A novel and efficient method utilizing Locked Nucleic Acids (LNA) to remove up to 99% rRNA | |
|---|---|---|
| RiboMinus Eukaryote Kit for RNA-Seq | RiboMinus Plant Kit for RNA-Seq | |
| Platform | Diagnostic | Diagnostic |
| Starting material | 1–10 µg total RNA 100–500 ng total RNA (low-input protocol) | 1–10 µg total RNA |
| RiboMinus RNA yield | 1–2 µg from 10 µg human total RNA ~100 ng from 500 ng human total RNA (low-input protocol) | 1–2 µg from 10 µg human total RNA |
| Probe specificity | Eukaryotic | Plant |
| Cat. No. | A1083708 - 8 preps | A1083808 - 8 preps |
RNA Library Construction
Library construction, template preparation, and sequencing are at the heart of the sequencing workflow. Utilization of sequencing controls and methods for data analysis complete the workflow and provide useful annotated data. Generate cDNA by reverse transcription from adaptors ligated to the ends of RNA, then amplify the cDNA using primers complementary to the adaptors, and purify it.
| Order now | Order now | Order now | |
| Prepare representative cDNA libraries for strand-specific RNA sequencing | A Great Solution for NGS | The Ultimate Solution for NGS | |
|---|---|---|---|
| Ion Total RNA-Seq Kit v2 | SOLiD Total RNA-Seq Kit | Ambion RNA-Seq Library Construction Kit | |
| Platform | Ion Torrent PGM | SOLiD 5500 Analyzer | Illumina |
| RNA input type | rRNA-depleted total RNA, poly(A) RNA, or miRNA | Total RNA, rRNA-depleted total RNA, poly(A) RNA, or miRNA | Total RNA, rRNA-depleted total RNA, poly(A) RNA, or miRNA |
| RNA input range | Small RNA: total RNA containing 1–100 ng of miRNA Whole transcriptome: 100 ng total RNA (~1 ng poly(A) or ~25 ng rRNA-RNA | Small RNA: total RNA containing 1–100 ng of miRNA Whole transcriptome: 100 ng poly(A), 200 ng | Small RNA: total RNA containing 1–100 ng of miRNA Whole transcriptome: 100 ng poly(A) or 200 ng rRNA-depleted total RNA, or 200 ng total RNA |
| Barcodes available | Yes | 96 | No |
| Cat. No. | 4475936 - 12 reaction kit | 4445374 - 12 reaction kit | 4454073 - 12 reaction kit |
RNA Library Controls
Library construction, template preparation, and sequencing are at the heart of the sequencing workflow. Utilization of sequencing controls and methods for data analysis complete the workflow and provide useful annotated data.
Learn more about ERCC Controls
| Order now | Order now | |
| 2 tubes containing preformulated mixes of the same 92 RNA transcripts but in different ratios, for measuring dynamic range, assessing bias, and confirming expression fold changes | Single tube containing a preformulated mix of 92 RNA transcripts for measuring dynamic range and assessing bias | |
|---|---|---|
| ERCC ExFold RNA Spike-In Mixes (2 x 10 µL) | ERCC RNA Spike-In Mix (1 x 10 µL) | |
| Top Seller | ||
| Compatible sample types | Eukaryotic total RNA, rRNA-depleted total RNA, or poly(A) RNA | Eukaryotic total RNA, rRNA-depleted total RNA, or poly(A) RNA |
| Dynamic range | 6 logs | 6 logs |
| Volume | 10 µL (2 tubes) | 10 µL |
| Number of reactions | Spike-in controls diluted based on sample type and concentration | Spike-in controls diluted based on sample type and concentration |
RNA Barcoding
Sets of 16 barcodes are available for use with the SOLiD Total RNA-Seq Kit or the SOLiD SAGE Kit with Barcoding Adaptor Module. Barcodes cost-effectively increase the number of samples that can be analyzed in a single sequencing run. Barcodes assign a unique identifier to templated beads made from a single library. Once assigned, the RNA libraries can be pooled and sequenced.
Cat. No. | Product |
RNA Sequencing
No matter which platform you choose to suit your needs, we continue to support the evolving needs of this field with gold-standard instrumentation, innovative next-generation technology, optimized best-in-class reagents, consumables, analysis software, applications and world-class technical support.
Next-Generation (SOLiD)
Sequence right here, right now—minimizing wait time and costs for wasted reagents.
Capillary Electrophoresis Sequencing
The gold standard in sequencing and a fast, flexible platform for analysis.
Semiconductor (Ion Torrent) Sequencing
Sequence from your benchtop with Ion Torrent revolutionary "on-chip" technology.
RNA & Transcriptome Sequencing
Choose the correct technology for your specific RNA application.
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Stylesheet for Classic Wide Template adjustments
仅供科研使用，不可用于诊断目的。