RNA reference guide
Sample preparation and purification solutions
Introduction
For over three decades and published in over 100,000 publications, scientists have trusted their most precious samples with innovative and robust RNA isolation and purification products from Thermo Fisher Scientific. Building on this legacy, scientists continue to develop new products and improve results with existing ones. Development of RNA-related research products began with Ambion in 1989. Ambion products are now offered by Thermo Fisher Scientific, integrating an innovative portfolio of RNA isolation products including organic reagents, columns, lysis buffers, and magnetic beads. Trusted products for RNA isolation include Applied Biosystems™ MagMAX™ kits, Invitrogen™ Cells-to-CT™, PureLink™, and mirVana™ kits, Invitrogen™ TRIzol™ Reagent, and Invitrogen™ Dynabeads™ isolation technologies. In biological research, RNA isolation is a crucial step in the journey to discovery. Whichever downstream application is being attempted, users can be confident in the portfolio of high-quality products and methods of extraction for all sample types.
Purification and Isolation Technologies
Thermo Fisher Scientific offers simple solutions to meet the complex needs of every experiment and research project. Purification methods range from organic extraction reagents to spin columns and magnetic beads.
Summary of RNA Purification Technologies
| Method | Organic Extraction | Spin Column | Crude Lysate | Magnetic Beads |
|---|---|---|---|---|
| Technology | Monophasic solution of phenol and guanidine isothiocyanate | Spin column and reagents optimized for RNA isolation via centrifugation | Lysis buffer and stop solution compatible with reverse transcriptase and PCR applications | Magnetic beads and reagents optimized for RNA isolation |
| Sample Type(s) | Multi-sample—see individual tabs for product details | Multi-sample—see individual tabs for product details | Cells in culture | Multi-sample—see individual tabs for product details |
| Technique | RNA isolation achieved through organic extraction and RNA precipitation | Homogenized samples loaded onto spin column; RNA is washed and eluted off column using benchtop centrifuge or vacuum manifold | Cells lysed with lysis buffer compatible with reverse transcriptase and qPCR master mix reactions; no RNA isolation required | Homogenized samples mixed with magnetic beads; beads are washed with wash buffers, and RNA is then eluted off the beads |
| Purity | Medium | High | Not applicable | Highest |
| Throughput | Low | Medium to high | Medium to high | Medium to high |
| Advantages | • Efficient lysis and isolation • Scalable format • Great for high-fat and cartilaginous samples • 30–60 min procedure |
• Easy to use • Processing of many sample types and volumes • Specialized equipment not required • High yield and purity • 20 min procedure • Isolates true total RNA, including miRNA |
• 7 min sample prep • Addition-only workflow—transfer to column or bead not required • Process 10–100,000 cells with single protocol • RNA isolation not required • Highest reproducibility and sensitivity • Complete kit contains lysis reagents, reverse transcriptase, and qPCR master mix • 1-step and 2-step workflows available |
• Higher-efficiency washes with bead movement • No column means no risk of clogging • Fully scalable sample prep to process low and high sample inputs • High yield and efficiency • High throughput-friendly • Automatable on magnetic particle handlers |
| Difficulties | • Novice users find phase separation challenging to master • Hazardous—chemical fume hood required • Challenging to process large number of samples simultaneously |
• Manual process • Processing individual columns often challenging to handle |
• Limited to cells in culture and blood | • Magnet stand required • Instrument required for automatic processing |
| Recommended for | Most sample types, but best for extraction of RNA from high-fat tissues (such as brain and mammary tissue) or infectious samples | High-throughput gene expression screening from cells in culture | Most RNA isolation needs cells in culture | Medium- to high-throughput or automatic sample prep processing |
Organic Extraction
Total RNA Isolation with TRIzol Reagent
Invitrogen™ TRIzol™ Reagent is a trusted and referenced reagent for isolating high-quality, intact RNA. It allows sequential precipitation of RNA, DNA, and protein from a single sample. After homogenizing the sample with TRIzol Reagent, chloroform is added, and the homogenate separates into a clear upper aqueous layer containing RNA, an interphase layer, and a pink lower organic layer containing DNA and protein. RNA is precipitated from the upper aqueous layer with isopropanol. DNA is precipitated from the interphase and organic layers with ethanol. Protein is precipitated from the phenol-ethanol supernatant with isopropanol. The precipitated RNA, DNA, or protein is washed to remove impurities and then resuspended for use in downstream applications.
Trust in TRIzol Reagent
Features of TRIzol Reagent:
- Unparalleled lysis capability
- Single-step protocol, monophasic solutions (phenol and guanidine isothiocyanate)
- Flexible formulations for difficult samples
- Low genomic DNA (gDNA) contamination of isolated RNA
- Can purify DNA, RNA, and protein all from the same sample
Figure 1. TRIzol Reagent is the most cited organic reagent for nucleic acid isolation, with citations gathered from 1990 to 2019 according to PubMed records (ncbi.nlm.nih.gov).
Did you know? The "TRI" in TRIzol stands for total RNA isolation. It also signifies the three layers that form during sample extraction, which allow RNA, DNA, and protein to be purified from a single source.
TRIzol Plus RNA Purification Kit
Figure 2 illustrates high yield of RNA using the Invitrogen™ TRIzol™ Plus RNA Purification Kit. This kit isolates more RNA faster by combining the lysis capability of TRIzol Reagent with the convenient RNA extraction technology of silica spin columns of the Invitrogen™ PureLink™ RNA Mini Kit.
Pro tip: The TRIzol Plus RNA Purification Kit combines TRIzol Reagent with spin column technology and can isolate up to 10 times more RNA than other purification systems.
TRIzol Reagent workflow:
Store blood samples with RNAlater Stabilization Solution ➡️ Isolate RNA in 1 hr with TRIzol Reagent ➡️ Generate first-strand cDNA with SuperScript IV First-Strand Synthesis System ➡️ Analyze gene expression using TaqMan Fast Advanced Master Mix.
Figure 3 shows safe isolation of RNA from blood samples for gene expression analysis. Invitrogen TRIzol and TRIzol LS Reagents have been demonstrated to chemically inactivate infectious viruses such as Ebola virus (EBOV) for improved safety. With TRIzol reagents, RNA purified from blood samples is ready for a variety of RNA analyses.
TRIzol Options Comparison
| Product | Product Size | Time | Sample Input | Final Product | Recommended For |
|---|---|---|---|---|---|
| TRIzol Reagent | 100 mL, 200 mL | N/A | Bacteria, Blood, Cells, Plant samples, Tissue (including fat and cartilage), Viral samples, Yeast | Total RNA, DNA, Protein | Gene expression, RT-qPCR, NGS, Microarray analysis, Cloning, Northern blotting, Nuclease protection assays, cDNA library construction |
| TRIzol LS Reagent | 100 mL, 200 mL | N/A | Blood, Liquid samples (e.g., serum), Viral samples | Total RNA, mRNA, miRNA, DNA, Protein | Gene expression, RT-qPCR, NGS, Microarray analysis, Cloning, Northern blotting, Nuclease protection assays, cDNA library construction |
| TRIzol Plus RNA Purification Kit | 50 preps | 1 hr | Bacteria, Blood, Cells, Plant samples, Tissue (including fat and cartilage), Viral samples, Yeast | Total RNA, mRNA, miRNA, DNA, Protein | Gene expression, RT-qPCR, NGS, Microarray analysis, Cloning, Northern blotting, Nuclease protection assays, cDNA library construction |
| TRIzol Max Bacterial RNA Isolation Kit | 100 preps, 200 preps | N/A | Bacteria | Total RNA, mRNA, miRNA, DNA, Protein | Gene expression, RT-qPCR, NGS, Microarray analysis, Cloning, Northern blotting, Nuclease protection assays, cDNA library construction |
Phasemaker Tubes
Invitrogen™ Phasemaker™ Tubes simplify TRIzol reagent use by creating a tight, durable seal between aqueous and organic phases, allowing for easy pipetting of the RNA phase. Benefits include higher yield (up to 30% increase), ease of use, ready-to-use convenience, and increased throughput.
Figure 4 illustrates how Phasemaker Tubes create a durable seal, allowing for maximized sample runs without losing phase separation and easy pipetting of the RNA phase with minimal loss.
Pro tip: Even if the tube is dropped after centrifugation, the TRIzol Reagent phases remain separated; do not waste time centrifuging again.
Ordering Information:
| Product | Quantity | Cat. No. |
|---|---|---|
| TRIzol Reagent | 100 mL, 200 mL | 15596026, 15596018 |
| TRIzol LS Reagent | 100 mL, 200 mL | 10296010, 10296028 |
| TRIzol Plus RNA Purification Kit | 50 preps | 12183555 |
| TRIzol Max Bacterial RNA Isolation Kit | 100 preps, 200 preps | 16096020, 16096040 |
| Phasemaker Tubes | 100 tubes | A33248 |
Column Purification
Spin Down for Flexible, Easy, and Reliable RNA Isolation
PureLink silica columns offer superior sample input capacity (6x more than other suppliers) and can process various sample types in 20 minutes. Kits are available for total RNA, mRNA, miRNA, small RNA, viral RNA, and sequence-specific RNA extractions.
A Tried and True Spin on RNA Purification
Samples are lysed using a guanidine isothiocyanate lysis buffer that liberates proteins and other debris from RNA while maintaining RNA integrity. Nuclease-free reagents and RNase-free silica membranes in spin columns enable a safe and easy procedure. After lysate passes through the silica membrane via centrifugation, the membrane is washed to remove contaminants. RNA is then eluted with RNase-free water and is ready for downstream applications.
Did you know? Most analytical methods for RNA isolation do not require the removal of contaminating gDNA. For qPCR applications, Invitrogen™ TaqMan® Gene Expression Assays can be used without additional DNase treatment. Save time and budget by using the right method.
If residual DNA removal is necessary, the PureLink RNA Mini Kit offers on-column treatment, or post-isolation DNase treatment with the Invitrogen™ PureLink™ DNase Set.
Pro tip: Combine TRIzol Reagent with spin column filter purification for exceptionally pure RNA.
Take a Spin with PureLink Purification Kits
PureLink RNA Mini Kit
Processes sample inputs from 5 to 200 mg of animal or plant tissue, or 0.5 x 10⁶ to 0.5 x 10⁸ tissue culture cells per spin column in 20 minutes. It isolates up to 1,000 µg of total RNA* with one column and offers very low residual DNA carryover. Flexible, optional on-column or post-isolation DNase treatment is available with the Invitrogen™ PureLink™ DNase Set.
Pro tip: The PureLink RNA Mini Kit works with various sample types including tissue, blood, bacteria, yeast, and plant cells, using optimized protocols.
Did you know? One PureLink RNA Mini Kit can process the same amount of sample as one Qiagen RNeasy™ Mini Kit and one RNeasy™ Midi Kit combined, offering broader sample input capacity.
A Closer Look
Figure 6 compares RNA yields from mouse liver, heart, and brain tissue. For all tissue types, the PureLink kit yielded more RNA than the Qiagen kit. For liver and brain tissue, the PureLink kit recovered approximately twice as much RNA as the Qiagen kit. Figure 6(D) shows the PureLink column has a larger bed than the Qiagen column, supporting higher input capacity.
Figure 7(A) compares purity measurements (A260/A280, A260/A230) for TRIzol Reagent, PureLink RNA Mini Kit, and TRIzol Plus kit. Figure 7(B) shows RNA integrity analysis using an Agilent Bioanalyzer™ instrument, indicating high RIN values and intact RNA were obtained.
Product Offerings
mirVana miRNA Isolation Kit: Isolates total RNA ranging from kilobases down to 10 bases, ideal for miRNA, siRNA, shRNA, and snRNA analysis. It offers efficient isolation of total RNA containing small RNA and a simple 30-minute procedure.
RecoverAll Multi-Sample RNA Isolation Kit for Special RNA Species: Recovers RNA and DNA from FFPE samples, maximizing RNA yield without degradation. A single spin column can tolerate large amounts of paraffin.
Pro tip: Combine TRIzol Reagent with spin-column filter purification for exceptionally pure RNA.
Did you know? Flexible, optional on-column or post-isolation DNase treatment is available with the PureLink DNase Set for easy removal of residual DNA.
Figure 8 illustrates an example workflow for protein expression analysis using RNA isolated with the PureLink RNA Mini Kit, ready for RT-PCR and downstream analysis.
Selection Guide for Column-Based RNA Purification Products
| Product | Isolation Technology | Product Size | Time | Sample Input | Final Product | Recommended For |
|---|---|---|---|---|---|---|
| PureLink RNA Mini Kit | Silica or glass fiber spin column | 10, 50, 250 preps | 20 min | Bacteria, Blood, Cells, Plant, Tissue, Yeast | Total RNA | Microarray analysis, NGS, Northern blotting, Nuclease protection assays, qPCR, RT-PCR, cDNA library construction |
| PureLink Viral RNA/DNA Mini Kit | Silica or glass fiber spin column | 50 preps | 45 min | Plasma, Serum, Cerebrospinal fluid, Cell-free fluids | gDNA | Cloning, PCR, qPCR, RT-PCR, Sequencing, NGS, Southern blotting, Northern blotting |
| mirVana miRNA Isolation Kit | Organic extraction and spin column | 40 preps | 30 min | Bacteria, Cells, Plant, Tissue, Virus, Yeast | Total RNA, transcriptome RNA, miRNA, siRNA, snRNA | miRNA analysis, NGS, Microarray analysis, qPCR, RT-PCR |
| RNAqueous Total RNA Isolation Kit | Silica or glass fiber spin column | 50 preps | 25–35 min | Bacteria, Cells, Plant, Tissue, Virus, Yeast | Total RNA | Microarray analysis, NGS, Northern blotting, Nuclease protection assays, qPCR, RT-PCR, cDNA library construction |
| RecoverAll Total Nucleic Acid Isolation Kit | Silica or glass fiber spin column | 40 preps | 75 min | FFPE and other fixed samples | Genomic DNA, total RNA, miRNA | NGS, Northern blotting, PCR, qPCR, RT-PCR, Southern blotting, cDNA library construction, miRNA analysis |
Ordering Information
See the table in the PDF for specific product ordering details (Cat. Nos. for mirVana miRNA Isolation Kit, PureLink RNA Micro Scale Kit, PureLink RNA Mini Kit, etc.).
Crude Lysate Purification
Cells-to-CT Values
Invitrogen™ Cells-to-CT™ kits enable high-throughput and high-sensitivity cell-based gene expression analysis in as little as 7 minutes without isolating RNA. These kits include cell lysis reagents, reverse transcriptases, and Applied Biosystems™ qPCR master mixes suitable for TaqMan Gene Expression Assays or SYBR™ Green PCR primer methods. 1-step and 2-step RT-qPCR and single-cell kits are available.
The Secret to Lysate-Based Gene Expression
The advanced Cells-to-CT Lysis Solution and Stop Solution are key to lysate-based gene expression analysis. The lysis buffer liberates RNA without interfering with downstream analysis.
Cells-to-CT Reverse Transcriptase and Master Mix Optimization
Thermo Fisher Scientific R&D scientists tested various reverse transcriptases and master mixes with Cells-to-CT lysis components to determine optimal combinations for RT-qPCR applications. Criteria included PCR efficiency, maximum crude lysate input, PCR inhibition, reproducibility, and linear amplification from 10 to 100,000 cells. Optimized reagents undergo rigorous quality control.
Newer Cells-to-CT kits feature faster and more sensitive reverse transcriptases and master mixes. The TaqMan Fast Advanced Cells-to-CT Kit processes a 96-well plate in 80 minutes, while the Invitrogen™ SYBR™ Green Fast Advanced Cells-to-CT Kit takes 90 minutes. Users report better sensitivity compared to first-generation kits.
Pro tip: For cost-conscious users, design custom primers and use the SYBR Green Fast Advanced Cells-to-CT Kit.
Did you know? The TaqMan Fast Advanced Cells-to-CT Kit was designed specifically for TaqMan Gene Expression Assays.
TaqMan Fast Advanced Cells-to-CT Kit
- No tedious RNA purification—no columns, heating, centrifugation, or sample transfer.
- Uses the same proprietary Lysis Solution and Stop Solution as the original TaqMan Gene Expression Cells-to-CT Kit.
- Extraordinary ease and speed—prepare 96 samples for RT-qPCR in typically under 10 minutes.
- Superior performance for consistent accuracy, reproducibility, and sensitivity with 10–100,000 cells.
- Premium validated kit workflows with pre-optimized reagents.
- Additional probe for superior specificity.
- Detects virtually any gene product with over 2 million predesigned assays.
- Affordable assays for human, mouse, and rat genes.
- TaqMan Assays available for 28 species and some pathogens.
SYBR Green Fast Advanced Cells-to-CT Kit
- No tedious RNA purification.
- Superior performance for consistent accuracy, reproducibility, and sensitivity with 10–100,000 cells.
- Uses the same proprietary Lysis Solution and Stop Solution as the original TaqMan Gene Expression Cells-to-CT Kit.
- Extraordinary ease and speed—96 samples for RT-qPCR in typically under 10 minutes.
- Premium validated kit workflows.
- High specificity for detecting targets.
Cells-to-CT Product Selection Guide
| PCR Method | Cells-to-CT 1-Step TaqMan Kit | Cells-to-CT 1-Step Power SYBR™ Green Kit | TaqMan Fast Advanced Cells-to-CT Kit | TaqMan Gene Expression Cells-to-CT Kit | SYBR Green Fast Advanced Cells-to-CT Kit |
|---|---|---|---|---|---|
| Specificity | High | Medium | High | Medium | High |
| Reproducibility | High | Medium | High | Medium | High |
| Sensitivity | 20% of lysate represented; 1–10 copies | 13.5% of lysate represented | 20% lysate | 20% lysate | 20% lysate |
| Product Size | 20, 100, 400 preps | 20, 100, 400 preps | 40, 100, 400 preps | 40, 100, 400 preps | 40, 100, 400 preps |
| Lysate Prep Time | 35 min | 90 min | 7 min prep | 110 min | 95 min |
| Total Workflow Time | N/A | N/A | 80 min | N/A | N/A |
| Sample Input | 10–100,000 cells | 10–100,000 cells | 10–100,000 cells | 10–100,000 cells | 10–100,000 cells |
| Final Product | RT-qPCR results | RT-qPCR results | RT-qPCR results; cDNA | RT-qPCR results; cDNA | RT-qPCR results; cDNA |
| Recommended For | Gene expression analysis using TaqMan Gene Expression Assays; Users who want faster time-to-results; Users who do not need to archive cDNA | Gene expression analysis using primers and SYBR Green assays; Users who want faster time-to-results; Users who do not need to archive cDNA | Gene expression analysis using TaqMan Gene Expression Assays; Users who want to archive cDNA; Users who need maximum sensitivity of detection | Gene expression analysis using TaqMan Gene Expression Assays; Users who want to archive cDNA | Gene expression analysis using primers and SYBR Green assays; Users who want to archive cDNA |
Magnetic Beads
Make Your Research Magnetic
Thermo Fisher Scientific offers a wide range of magnetic bead platforms, including Applied Biosystems™ MagMAX™ reagents and Invitrogen™ Dynabeads™ magnetic beads. These products balance high yield and reproducibility with low nonspecific binding and are compatible with Thermo Scientific™ KingFisher™ instruments for high-throughput applications.
MagMAX Magnetic Beads
MagMAX™ magnetic bead technology utilizes uniform, mono-sized superparamagnetic beads paired with an improved elution buffer for high binding efficiency and reproducibility. MagMAX kits simplify nucleic acid extraction, making it sensitive, rapid, and automation-friendly.
MagMAX mirVana Total RNA Isolation Kit
This flexible magnetic bead-based kit isolates total RNA from kilobases down to 10 bases. It can elute RNA in as little as 20 µL of nuclease-free water for more concentrated RNA. The kit purifies 6 to 96 samples at once and is automatable with Thermo Scientific™ KingFisher™ magnetic particle processors.
- Binds RNA more efficiently than silica-membrane methods.
- Higher, more consistent RNA yields.
- Streamlined protocols for phenol-free, scalable isolation of total RNA.
- Recovers pure miRNA compatible with miRNA-Seq and RT-qPCR methods.
Figure 11 compares MagMAX mirVana with the Invitrogen™ mirVana™ PARIS™ kit, showing greater sensitivity for miRNA targets using the MagMAX mirVana kit.
Did you know? MagMAX kits were launched in 2004 for animal health testing, providing fast, affordable RNA extraction.
MagMAX Plant RNA Isolation Kit
Designed to isolate total RNA from various plant species and tissue types, including woody, polyphenol-rich, and lignified samples. It provides high yields of pure total RNA, isolates high-quality intact RNA free of inhibitors and gDNA contamination, and is adjustable for high-throughput automation.
Dynabeads Magnetic Beads
The Invitrogen™ Dynabeads™ mRNA DIRECT™ Kit is designed for simple and rapid isolation of pure, intact polyadenylated (poly(A)) mRNA directly from crude lysates of animal and plant cells and tissues. It offers a flexible protocol scalable for all sample sizes and can isolate mRNA from mammalian, fish, amphibian, insect, and plant origins. High capture efficiency facilitates detection by RT-PCR of mRNA from specialized cells.
Figure 12 illustrates the Dynabeads mRNA DIRECT Kit protocol, involving binding of poly(A) RNA to oligo(dT)25 coated beads, washing, elution, and reverse transcription.
Magnetic Bead-Based Purification Product Selection Guide
| Product | Product Size | Time | Sample Input | Final Product | Recommended For |
|---|---|---|---|---|---|
| MagMAX mirVana Total RNA | 6–96 reactions | 1.5–2 hr | Blood cells, Liquid samples (e.g., serum), Tissue | Total RNA* | NGS, qPCR, miRNA analysis |
| Dynabeads mRNA DIRECT Purification Kit | 20 preps with 5 mL, 40 preps with 10 mL | 15 min | Blood cells, FFPE and other fixed samples, Liquid samples (e.g., serum), Plant samples, Tissue, Viral samples, Yeast | mRNA | Cloning, Northern blotting, Nuclease protection assays, qPCR, RT-PCR, cDNA library construction |
| MagMAX Viral/Pathogen Ultra Nucleic Acid Isolation Kit | 100 preps | <60 min | Serum, Plasma, Urine, Cerebrospinal fluid (CSF), Universal viral transport media, Whole blood, Bronchoalveolar lavage (BAL), Swabs, Toenail fungus | RNA and DNA | NGS, Real-time PCR |
| MagMAX Cell-Free Total Nucleic Acid Isolation Kit | 50 samples for 2 mL plasma input | 90 min | Serum, Plasma, Urine | cfRNA and cfDNA | NGS, qPCR |
| MagMAX Microbiome Ultra Nucleic Acid Isolation Kit | 25 samples for 4 mL plasma input, 100 preps | 60 min | Stool, Soil, Swabs | RNA and DNA | Microarrays, qPCR, NGS |
| MagMAX FFPE DNA/RNA Ultra Kit | 96 isolations (48 RNA and 48 DNA, or 96 DNA, or 96 RNA) | 4 hr total time (1 hr hands-on time) | FFPE tissue samples, such as resections, biopsies, and aspirates | RNA and DNA | NGS, PCR, RT-PCR (endpoint), qPCR, miRNA analysis |
| MagMAX Plant RNA Isolation Kit | 384 preps | 75 min | Any plant sample | Total RNA | RT-PCR (endpoint), qPCR, Sequencing, Northern blotting, Nuclease protection assays |
Instruments
KingFisher Instruments
Pair MagMAX purification kits with KingFisher instruments to automate RNA extractions. KingFisher instruments offer easy-to-follow protocols for RNA isolation from most sample types, saving time and reducing user error. They automate DNA, RNA, protein, and cell extraction by moving magnetic beads, ensuring clean extractions and consistent results through a 4-step process: lyse, bind, wash, and elute.
KingFisher Instrument Comparison
| KingFisher Instrument | Duo Prime | Flex | Presto |
|---|---|---|---|
| Instrument Size | Compact benchtop | Benchtop | Benchtop—integrates with robotic liquid handler |
| Throughput Level | Low to medium | Medium to high | Ultrahigh |
| Processing Volume Range | 50–1,000 µL; 200–5,000 µL | 20–100 µL; 20–200 µL; 50–1,000 µL; 200–5,000 µL | 50–1,000 µL; 200–5,000 µL; 50–150 µL |
| Samples per Run | 6 or 12 | 24 or 96 | 24 or 96 |
| Customizable Protocols | Yes, with PC software | Yes, with PC software | Yes, with PC software |
| Heating/Cooling | 10°C to 75°C; 4°C to 75°C | From 5°C above ambient temperature to 115°C | From 5°C above ambient temperature to 115°C |
| Ultraviolet Lamp | 8 watts (up to 16 hr) | No | No |
Qubit Flex Fluorometer
The Invitrogen™ Qubit™ Flex Fluorometer is a benchtop fluorometer for accurate quantification of DNA, RNA, miRNA, and protein. It measures up to 8 samples simultaneously, reducing assay variability. Paired with Qubit reagents, it provides highly accurate and reproducible results.
- Measures up to 8 samples, quantifying DNA, RNA, and protein up to 50% faster than single-sample readers.
- Works seamlessly with Qubit reagents for reliable, sensitive, and specific results.
- Highly accurate measurements using only 1–20 µL of sample, even with dilute samples.
- Quickly generates working solution preparation instructions and determines sample molarity.
Figure 14 shows time savings with the Qubit Flex Fluorometer, demonstrating up to 50% time savings when measuring more samples.
NanoDrop One Spectrophotometers
Thermo Scientific™ NanoDrop™ One and OneC Microvolume UV-Vis Spectrophotometers check DNA, RNA, and protein quantity and quality using only 1–2 µL of sample in seconds. Thermo Scientific™ Acclaro™ sample intelligence technology provides a deeper understanding of sample quality.
- Achieve accurate results with full spectral data.
- Identify sample contaminants (phenol, guanidinium salts, protein) with sophisticated algorithms.
- Easily transfer data to PC or network via Wi-Fi, Ethernet, or USB.
- Print results from on-board control.
- Access data anywhere, anytime with Connect cloud platform.
Figure 15 shows purity analysis of RNA from three different tissue types using the NanoDrop spectrophotometer, indicating high RNA purity with A260/A280 ratios between 2.1 and 2.2.
Tips and Tricks for Specialty Samples
Sample Types
The biological nature of your sample type influences the purification method chosen. Mammalian cells, which have easily broken membranes, can be purified by various methods; crude lysates may suffice for RT-qPCR, while NGS requires higher purity. Brain and kidney tissues, high in fat, benefit from TRIzol Reagent combined with Phasemaker Tubes and PureLink columns. Blood cells are heterogeneous and may require specialty kits for sensitive applications like NGS; PureLink Mini Kits are suitable for other applications. Bacteria, yeast, and plants have different cell walls requiring specific reagents, with specialty kits available.
Specialty Kits for RNA Purification
| Sample Type | Special Stabilization | 1–24 Samples | >25 Samples |
|---|---|---|---|
| Cultured cells | PureLink RNA Mini Kit (p 12) TRIzol Reagent (p 7) |
Cells-to-CT kits (p 21) PureLink Pro 96 Total RNA Purification Kit (p 16) |
|
| Blood | RNAlater Solution | PureLink RNA Mini Kit (p 12) TRIzol LS Reagent (p 9) |
MagMAX mirVana Total RNA Isolation Kit (p 23) Dynabeads mRNA DIRECT Kit (p 24) |
| Fresh or frozen brain and kidney tissue | RNAlater Solution | TRIzol Plus RNA Purification Kit & Phasemaker Tubes Complete System (p 10) MagMAX mirVana Total RNA Isolation Kit (p 23) |
PureLink RNA Mini Kit (p 12) |
| Fresh or frozen other animal tissues | RNAlater Solution | PureLink RNA Mini Kit (p 12) TRIzol Plus RNA Purification Kit & Phasemaker Tubes Complete System (p 10) |
MagMAX mirVana Total RNA Isolation Kit (p 23) |
| FFPE samples | RecoverAll Total Nucleic Acid Isolation Kit for FFPE (p 14) | MagMAX FFPE DNA/RNA Ultra Kit (p 25) | |
| Plant | PureLink RNA Mini Kit (p 12) PureLink Plant RNA Reagent (p 12) |
MagMAX Plant Isolation Kit (p 24) | |
| Yeast | PureLink RNA Mini Kit (p 12) | Dynabeads mRNA DIRECT Purification Kit (p 24) MagMAX Viral/Pathogen Ultra Nucleic Acid Isolation Kit (p 25) |
|
| Bacteria | PureLink RNA Mini Kit (p 12) TRIzol Max Bacterial RNA Isolation Kit (p 9) |
MagMAX Total Nucleic Acid Isolation Kit | |
| Virus | PureLink Viral RNA/DNA Mini Kit (p 16) RNAqueous Total RNA Isolation Kit (p 16) |
MagMAX Viral/Pathogen Ultra Nucleic Acid Isolation Kit (p 25) |
Sample Prep Essentials
RNA is sensitive to degradation by RNases. Essential products to avoid, detect, and inhibit RNases include:
Invitrogen™ RNAlater™ Solution for RNA Storage and Stabilization
This room temperature-stable solution allows RNA storage without liquid nitrogen. Thin tissue samples (0.5 cm) ensure quick permeation before RNases destroy RNA. Remove tissue from solution and suspend as if freshly harvested.
- Aqueous, nontoxic tissue storage reagent.
- Eliminates RNases and rapidly permeates tissue to stabilize and protect cellular RNA.
Invitrogen™ Nuclease-Free Water
Purified water ensures consistency when preparing reagents and resuspending RNA. Thoroughly rinse lab surfaces to reduce inhibitory residues and contaminants.
- Endonuclease-, exonuclease-, DNase-, and RNase-free water.
- Filtered, autoclaved, and rigorously tested according to ISO 9001 specifications.
- DEPC-treated and non-DEPC-treated options available.
RNaseZap Solution for Surface Decontamination
This room temperature-stable solution is for quick and easy use. RNaseZap wipes decontaminate pipettes and hard-to-reach surfaces. It helps ensure labware, instruments, and surfaces are RNase-free.
Invitrogen RNase-Free Tubes
Available in PCR, microcentrifuge, and conical tube formats. They are autoclave-safe, nuclease-free, and have undergone rigorous testing.
SUPERase-In RNase Inhibitor for RNA Inhibition
Effective at pH 5.5–8.5 and active up to 65°C. It removes high concentrations of dried-on RNase A and can be added to solutions to prevent RNase contamination. It inhibits RNases A, B, C, 1, and T1 without interfering with other enzymes like RNA polymerases.
Ordering Information: See the PDF for ordering details on RNA lab essentials, including RNase-Free Tips, PCR Tubes, Microfuge Tubes, RNAlater Stabilization Solution, and more.
Services and Support
Thermo Fisher Scientific provides fast responses from experienced scientists. Over 1,300 service and support specialists worldwide offer:
- Service plans: Planned maintenance and guaranteed response times to minimize downtime and extend instrument life.
- Compliance services: Timely, cost-effective, and audit-ready documentation for instrument installation and operation.
- Analytical validation (AV) consulting services: Technical project management and data analysis support for assay validation workflows.
- Bioinformatics and IT services: Consulting with bioinformatics application scientists for software, applications, and workflow optimization.
- Education services: Application and instrument training programs offered globally.
Find out more about services and support at thermofisher.com/instrumentservices.
RNA Technical Resources
Thermo Fisher Scientific is an essential resource for nucleic acid purification and analysis support. Navigate categories for technical information, tips, and answers to everyday problems.
Support
- thermofisher.com/napsupport
- thermofisher.com/technicalresources
- Email: techsupport@thermofisher.com
Web Resources
- thermofisher.com/contactus
- thermofisher.com/prepforsuccess
- thermofisher.com/rnaextraction
- thermofisher.com/magmax
- thermofisher.com/kingfisher
- thermofisher.com/rnaessentials
Find out more and order at thermofisher.com/rnabasics.
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