RNA extraction products include kits and reagents for purification of total RNA, transcriptome RNA, mRNA, microRNA and other small RNAs, and sequence-specific RNA. We also offer purified RNA. Our comprehensive products provide research scientists with the quality and performance they need. Our full suite of products is backed by scientists and engineers and is designed for RNA isolation and purification from a wide range of different sample types, throughputs, and input quantities.
Total RNA Extraction
Starting with high quality, pure, and intact total RNA is critical to many experiments, (e.g. RT-PCR, qRT-PCR, array analysis, Northern blots, nuclease protection assays and RNA sequencing). The table below will help you choose the right Invitrogen product to purify total RNA from your specific sample type and sample size.
Which Total RNA Purification Kit is right for you?
Order now | Order now | Order now | Order now | |
Gold standard for highly pure, intact RNA | High-quality RNA in less than 20 minutes | High-throughput purification of RNA and DNA | Complete, no purification system for qRT-PCR results | |
---|---|---|---|---|
TRIzol Reagents | PureLink Kits | MagMAX Kits | Cells to Ct | |
Recommendations | Most Cited, Top Seller | Best for qPCR
| ||
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Scalable, flexible format with magnetic beads | Chemical lysis (time to qRT-PCR results) |
Prep time | ~1 hr | < 20 minutes | ~45 minutes | 10 minutes
|
Compatible sample types | Most sample types, particularly more difficult to lyse | Cells and tissues | Tissues, cells, blood, serum/plasma, urine, plants | Cells
|
Amount of starting material | 100 mg of tissue or 107 cells (requires 1 mL reagent) | Up to 200 mg tissue, 5x106 to 1x108 cells | Up to 100 mg tissue, Up to 5x106 cells | 1–100,000 cells
|
Reverse transcription reagents included | | |||
qPCR Master mix included | ||||
High throughput compatible | (plate format available) |
Organic extraction & purification
Organic solvent extraction offers tried-and-true performance, and is ideal for the most stringent applications when combined with silica-membrane filtration or magnetic bead purification. Invitrogen TRIzol Reagent offers:
- Broadest sample type compatibility
- Robust method
- Good for difficult samples (e.g. high lipid tissue) and scalable sample input
- Wide acceptance as the gold standard
Magnetic bead purification
Magnetic particles offer solution-phase binding and washing kinetics to facilitate RNA purification and improved sample handling, including automation, ideal for higher throughput processing. Applied Biosystems MagMAX Magnetic Beads offer:
- Flexible and scalable throughput
- Amenable to automation for higher throughput
- Better removal of inhibitors and low elution volume resulting in more concentrated RNA
Invitrogen RNA essentials
Invitrogen products, are designed to minimize nuclease contamination and ensure experimental success in less-than-ideal conditions. These products are manufactured under the most stringent quality specifications and include:
- RNA purification kits
- Specimen collection and RNA stabilization solutions
- Nuclease inhibitors and decontaminants
- Nuclease-free buffers and plastics
- RNA ladders
- Northern blot reagents
Silica-membrane purification
Silica-membrane filters allow fast and easy RNA purification resulting in pure RNA that can be used in most applications. Invitrogen PureLink purification columns offer:
- A fast and easy method suitable for most downstream uses
- Wide sample type compatibility
- Familiar spin column format commonly used for low-med throughput
Total Nucleic Acid Extraction / Protein
Research sometimes requires testing of more than just RNA from each sample. For example researchers may want to compare the effects of genetic variations on gene expression, or to evaluate both protein and cognate mRNA levels. To serve this need, some Ambion purification products are designed to extract both RNA and DNA, or both RNA and protein from samples. Kits with these capabilities allow users to get more out of their samples while employing one kit to streamline the process.
The MELT Total Nucleic Acid Isolation Kit provides a method to obtain RNA or DNA from fresh or frozen tissue without physical digestion. For FFPE samples, the RecoverALL Total Nucleic Acid Isolation Kit for FFPE sets the standard and allows users to purify both RNA and DNA via a conventional filter-based method, while the new MagMAX FFPE Total Nucleic Acid Isolation Kit offers the same RNA and DNA purification from FFPE samples in a magnetic bead format without the need for xylene.
Nucleic Acid & Protein Extraction Kits
The PARIS Kit is designed to simultaneously purify total RNA and generate a native lysate suitable for common protein analysis techniques. The related mirVana PARIS Kit can also be used to obtain both total RNA and native lysate, but it also recovers the small RNA fraction, either along with total RNA or as a fraction enriched for small RNAs.
Total Nucleic Acid Extraction Kits
The MagMAX Total Nucleic Acid Isolation Kit purifies both RNA and genomic DNA with a combined mechanical (zirconia-bead beating) and chemical lysis method designed specifically for hard-to-lyse bacteria. Conversely, the MELT Total Nucleic Acid Isolation Kit provides a closed-tube, enzymatic method to obtain RNA or DNA from fresh or frozen tissue without physical disruption. For FFPE samples, the RecoverALL Total Nucleic Acid Isolation Kit for FFPE is designed to purify both RNA and DNA via a conventional glass fiber filter-based method, while the new MagMAX FFPE Total Nucleic Acid Isolation Kit offers xylene-free RNA and DNA purification from FFPE samples using a magnetic bead method.
Publication
MELT Total Nucleic Acid Isolation System: a new technology for hands-free tissue disruption, RNA preservation and total nucleic acid purification. Nature Methods, 23 Aug 2005.
microRNA & small RNA Isolation
MicroRNAs (miRNAs) and small interfering RNAs (siRNAs) modulate gene expression. Along with several other classes of small noncoding RNAs (e.g. snRNA, snoRNA and piRNA), they have tremendous impact on a variety of biological processes.
While most traditional RNA isolation methods are not very efficient at recovering these smaller RNAs, several kits developed by our RNA experts offer quantitative recovery of small RNAs from a variety of sample types.
Which microRNA & small RNA isolation kit is right for you?
Order now | Order now | Order now | Order now | |
30 min isolation from most samples | Recover all RNAs as well as proteins | HTP isolation of total RNA enriched for small RNAs | Complete kit, sample-to-qPCR from cells | |
---|---|---|---|---|
mirVana miRNA Isolation Kit (with or without phenol) | mirVana Paris | MagMAX mirVana Total RNA Isolation Kit | TaqMan microRNA Cells-to-CT Kit | |
Top Seller | ||||
RNA types isolated | Small & large RNA molecules | Small & large RNA molecules | Small & large RNA molecules | Small & large RNA molecules |
Top Seller | ||||
Prep time | 20 minutes | 30 minutes | <1 hour | 10 minutes |
Compatible sample types | Most sample types | Most sample types | Serum/Plasma, whole blood, Fresh/frozen tissue, cells, urine | Cells |
Isolation method | Highest purity and convenience; includes both organic extraction and silica column | Highest purity and convenience; includes both organic extraction and silica column | Scalable, flexible format with magnetic beads | Cells-to-CT direct lysis, no purification required |
Reverse transcription reagents included | No | No | No | TaqMan MicroRNA Reverse Transcription Kit |
qPCR Master mix included | No | No | No | TaqMan |
Kit size | 40 preps | 40 preps | 96 preps | 100 lysis reactions - plus - reagents for 200 qRT-PCR assays |
Publications
- Johnson CD, Esquela-Kerscher A, Stefani G, Byrom M, Kelnar K, Ovcharenko D, Wilson M, Wang X, Shelton J, Shingara J, Chin L, Brown D, Slack FJ. The let-7 MicroRNA Represses Cell Proliferation Pathways in Human Cells. Cancer Res. 2007 Aug 15;67(16):7713-22. [Abstract]
- Shingara J, Keiger K, Shelton J, Laosinchai-Wolf W, Powers P, Conrad R, Brown D, Labourier E. An optimized isolation and labeling platform for accurate microRNA expression profiling.RNA. 2005 Jul 25; [Epub ahead of print] [Abstract]
- Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, Labourier E, Reinert KL, Brown D, Slack FJ. RAS is regulated by the let-7 microRNA family. Cell. 2005 Mar 11;120(5):635-47. [Abstract]
- Cheng AM, Byrom MW, Shelton J, Ford LP. Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res. 2005 Mar 1;33(4):1290-7. [Abstract]
Technical Resources
- Potent Reagents for miRNA Studies
- Getting Started with MicroRNA Research
- MicroRNA Cloning Overview
- Prepare siRNA and miRNA Probes in Just 1 Hour
- Tips from the Bench: The Optimal Thickness of FFPE Sections for miRNA Isolation
- Isolate and Quantitate miRNA from Plants
- Tips from the Bench: High Throughput miRNA Isolation
- Tips from the Bench: Stability of miRNA at Sub-Zero Temperatures
- Your Data: Isolation of miRNAs from Pancreatic Samples
- Isolating miRNA for Profiling Studies
- Extract microRNA from Blood Samples
- Examine microRNA Profiles from Archived, Formalin-fixed, Paraffin-embedded (FFPE) Tissues
- Got Small RNA?
- Get the miRNA Out—Plant Applications
- Definition and Overview
- miRNA Function
- Expression & Targets
- miRNA Processing
mRNA Extraction
Which mRNA purification kit is right for you?
Order now | Order now | Order now | Order now | |
Rapid mRNA purified from total RNA | No purification needed, mRNA direct from crude samples | mRNA from micro-sized samples Best for next-generation sequencing of mRNA | HTP mRNA directly or enrich from total RNA | |
---|---|---|---|---|
Dynabeads\ mRNA Purification Kit | Dynabeads mRNA DIRECT | Dynabeads mRNA DIRECT Micro Kit | mRNA Catcher Plus | |
Top seller | ||||
Prep time | 15 minutes | 15 minutes | 15 minutes | 1.5 hours |
Direct isolation of mRNA from crude samples | No | Yes | Yes | Yes |
mRNA enrichment from total RNA | Yes | No | No | Yes |
Amount of starting material | <75 μg Total RNA | <20 x 106 cells 2–200 mg animal tissue 4–400 mg plant tissue | <1 x 104 cells <5 mg plant or animal tissue | 100 to 1 x 106 cells <4 mg tissue 40 µL whole blood 100 ng–100 µg total RNA |
Format | Fast magnetic bead capture from pure total RNA | Fast magnetic bead capture directly from crude samples | Fast magnetic bead capture directly from micro-size samples | HTP ready 96-well coated plate |
High-throughput compatible | Yes | Yes | Yes | Yes |
Kit size | 2 mL oligo dT beads | 5 mL or 10 mL oligo dT beads | 2 mL oligo dT beads (sufficient for 100 mRNA preps) | 96 or 960 preps |
Magnetic Bead mRNA Extraction and Enrichment Kits
Oligo(dT)/Magnetic Capture Methods
Column methods for mRNA purification are reliable but require extensive manual interaction, which means more time and reduced throughput. Magnetic beads conjugated with oligo(dT) offer additional handling and scaling benefits; however, this procedure does require the use of magnetic stands.
The Invitrogen Ambion Poly(A)Purist MAG Kit employs the same hybridization and wash solutions as those in the Poly(A)Purist Kit but with the added ease, speed, and scalability of magnetic beads. The protocol typically takes only 45 minutes, and the kit performs from 8 large enrichment reactions (up to 1 mg total RNA input) to 80 small enrichment reactions (as little as 30 µg total RNA input). Like the Poly(A)Purist Kit, this kit only enriches mRNA from previously purified total RNA.
The Invitrogen Dynabeads mRNA Purification Kit for mRNA Purification from Total RNA Preps, enriches mRNA from purified total RNA, typically in 15 minutes, and allows users the flexibility to elute the mRNA from the beads in as little as 5 µL, or even to perform cDNA synthesis directly on the beads without elution.
For those who prefer using Dynabeads technology with sample lysis and enrichment in one kit, the Invitrogen Dynabeads mRNA DIRECT Kits enable 15-minute lysis and enrichment processing of a variety of sample types. In fact, it is possible to capture mRNA and generate cDNA libraries from a single cell using this technology. The flexible format allows users to scale up or down and gives the option to elute the mRNA from the beads in as little as 5 µL, or to perform cDNA synthesis directly on the beads without elution. The 5 mL size is sufficient for 20 standard isolations, while the 10 mL size is sufficient for 40 standard isolations.
For those who work with smaller inputs, the Invitrogen Dynabeads mRNA DIRECT Micro Kit provides enough reagents for 100 mRNA isolations from up to 2.5 x 104 mononuclear cells, up to 1 x 104 cultured cells, or up to 5 mg tissue (depending on the tissue), with the same elution flexibility as the other Dynabeads kits.
Bacteria lack the relatively stable poly(A) tails found on eukaryotic mRNA. Until very recently, isolating mRNA from bacteria has been virtually impossible. The Invitrogen MICROBExpress Bacterial mRNA Isolation Kit employs a novel technology to remove up to >95% of the 16S and 23S rRNA from total RNA of E. coli and other bacterial species. The kit is suitable for mRNA enrichment from ≤10 µg of purified bacterial total RNA in about 2 hours from a broad spectrum of gram-positive and gram-negative bacteria. mRNA isolated with the MICROBExpress Kit is a superior template for synthesizing labeled cDNA for array analysis and is ideal for quantitative RT-PCR, northern blotting, and cDNA library construction.
mRNA Extraction and Enrichment via Coated Plates
The most distinctive mRNA purification method takes advantage of specially treated plates in conjunction with optimized reagents to both lyse and enrich mRNA from cells, tissue, blood, or purified total RNA. The Invitrogen mRNA Catcher PLUS product utilizes a proprietary surface treatment of a 96-well plate to facilitate easy and automation-friendly mRNA purification. Locked Nucleic Acid (LNA) technology incorporated into the 20-mer oligo(dT) increases specificity and hybridization efficiency while removing centrifugation, precipitation, and phase-separation steps from the overall workflow.
The mRNA Catcher PLUS Kit is capable of handling 100 ng to 100 µg total RNA, 100 to 106 mammalian cells, up to 4 mg tissue, and 40 µL whole blood. The binding capacity is >80 ng/well. The protocol employs a simple lysis, hybridization, wash, and elution process that can be completely automated on open-format liquid handlers in approximately 1.5 hours from start to finish. The system also has the flexibility to elute mRNA or to perform cDNA synthesis directly in the hybridization plate without elution.
Sequence-Specific RNA Extraction
Target a specific RNA sequence and isolate only those RNA molecules directly from crude lysates or other biological fluids using Dynabeads. Captured RNA is suitable for a wide range of downstream applications, including Dynabeads Streptavidin are ideal for numerous applications, including purification of proteins, nucleic acids purification, protein interaction studies, immunoprecipitation, immunoassays, phage display, biopanning, drug screening and cell isolation.
Which Sequence-Specific RNA Purification Kit or Reagent is Right for You?
Order now | Order now | Order now | |
4 beads in one kit, identify best for your application | Best solution for viscous samples | High surface area for maximum enrichment | |
---|---|---|---|
Dynabeads Streptavidin Trial Kit | Dynabeads M-270 Streptavidin | Dynabeads MyOne Streptavidin C1 | |
Top seller | |||
Bead size | 1 µm x 2, 2.8 µm x 2 | 2.8 µm | 1 µm |
Biotinylated Ig (µg/mg beads) | up to 20 | up to 10 | up to 20 |
Binding Capacity (Free Biotin) | 6502,500 pmoles/mg beads | 650–1,350 pmoles/mg beads | 2,500 pmoles/mg beads |
Pack size | 4 x 1 mL | 2 or 10 mL | 2 or 10 mL |
Ligand type | Streptavidin | Streptavidin | Streptavidin |
Dynabeads Technology
Our Streptavidin-coupled Dynabeads are a robust and versatile tool that can be used to literally target and capture specific RNA or DNA sequences and then pull them directly out of solution. These monosized superparamagnetic Dynabeads provide an efficient and solid-phase alternative to nitrocellulose and provide you with an unmatched level of product quality and data consistency. Excellent near liquid phase reaction kinetics allow for extremely fast protocols. The inherent ease of magnetic handling mean that downstream manipulations and buffer changes are as simple as concentrating the bead-bound target at the tube-wall with a magnet and then discarding the supernatant. These beads are compatible with an extremely broad range of sample types including most bodily fluids, crude lysates of plant, animal and microbial origin as well as purified total RNA or DNA. Since these Dynabeads will only interact with specifically targeted RNA or DNA molecules, upstream purification of total RNA or DNA is almost always an unnecessary step.
The 1 µm Dynabeads MyOne Streptavidin C1 present a very high surface area per mg of beads, enabling high enrichment of low abundance RNA or DNA. When the goal is to capture nucleic acid from more viscous samples such as cerebrospinal fluid, the larger 2.8 µm sized Dynabeads M-270 Streptavidin are recommended. These Dynabeads (MyOne Streptavidin C1 and M-270 Streptavidin) are optimally designed to have slightly negatively charged surfaces which ensure negligible non-specific binding of non-target nucleic acid sequences.
Examples of applications include; isolation of RNA/DNA infectious agent (1,2,3,4), subtractive hybridization (5,6,7), cDNA selection and enrichment, detection and isolation of mutated sequences (8,9,10), isolation of cell specific transcripts and mRNA differential display.
Learn more about:
The direct capture procedure involves the immobilization of double-stranded PCR products onto the beads. These are easily converted to single-stranded bead-bound templates which are then used to capture specific RNA or DNA molecules directly from solution.
An alternative indirect capture approach will offer faster reaction kinetics in some cases. This indirect capture procedure allows the target sequence to be captured prior to being immobilized the magnetic beads. First, a biotinylated capture-sequence (single-stranded DNA) is incubated with the sample and allowed to hybridize to the targeted RNA or DNA molecules in solution. Streptavidin coated Dynabeads are then added to the mixture and the hybridized sequences are immobilized onto the Dynabeads via the streptavidin-biotin bond.
Did you know?
Did you know that Dynabeads are employed on more than 25,000 routine IVD instruments worldwide?
Selected References
- Meng Q. et al. (2001) Automated multiplex assay system for simultaneous detection of hepatitis B virus DNA, hepatitis C virus RNA and human immunodeficiency virus type 1 RNA. J.Clin.Microbiol. 39(8):2937-2945.
- Stevens SJC. et al. (1999) Monitoring of Epstein-Barr virus DNA load in peripheral blood by quantitative competitive PCR. J.Clin. Microbiol. 37:2852-2857.
- Mangiapan G. et al. (1996) Sequence capture-PCR improves detection of mycobacterial DNA in clinical specimens. J. Clin. Microbiol. 34(5):1209-1215.
- Shuber AP. et.al. (2002) Accurate, noninvasive detection of Helicobacter pylori DNA from stool samples: potential usefulness for monitoring treatment. J. Clin. Microbiol. 40(1):262-264.
- Hansen-Hagge TE. et.al. (2001) Identification of sample-specific sequences in mammalian cDNA and genomic DNA by the novel ligation-mediated subtraction (LIMES). Nucl. Acids Res. 29(4):e20.
- Pradel N. et.al. (2002) Genomic subtraction to identify and characterize sequences of Shiga toxin-producing Escherichia coli O91:H21. Appl. Env. Microbiol. 68(5):2316-2325.
- Laveder P. et.al. (2002) A two-step strategy for constructing specifically self-subtracted cDNA libraries. Nucleic Acids Res. 30(9):e38.
- Lindblad-Toh K.et.al. (2000) Large-scale discovery and genotyping of single-nucleotide polymorphisms in the mouse. Nature Genetics. 24:381-386.
- Miyashiro I. et.al. (2001) Molecular strategy for detecting metastatic cancers with use of multiple tumor-specific MAGE-A genes. Clin.Chem. 47(3):505-512.
- Dong SM. et.al. (2001) Detection of colorectal cancer in stool with the use of multiple genetic targets. J Natl. Cancer Inst. 93(11):858-865.
Purified RNA
Ambion RNA experts are committed to providing the highest-quality prepared RNA available. To achieve this goal we are continually striving to understand what "high-quality RNA" truly means with respect to various downstream applications. Intactness of mRNA, genomic DNA contamination, presence/absence of enzymatic inhibitors, and ribosomal RNA contamination in poly(A) preparations are all important aspects of RNA quality. As we continue to gain a better understanding of how these factors affect downstream applications, we will strive to ensure that our FirstChoice RNA products exceed the requirements of even the most stringent applications.
- Certified to contain small RNAs (miRNA, siRNA, and snRNA)
- RNA available from a wide variety of human tissues and cell lines
- DNase-treated to remove contaminating DNA
- Superior quality control standards
Total RNA
We offer FirstChoice total RNA from human, mouse, and rat tissues, as well as from E. coli and human cell lines. RNA is obtained using Ambion RNA isolation reagents to yield highly pure, intact RNA that includes the small-RNA fraction. Purified RNA is subjected to a stringent DNase treatment. The RNA is supplied ready to use in any downstream application, including RT-PCR. Note that although it is impossible to remove every DNA molecule from an RNA sample, each preparation is tested to ensure that any residual DNA contamination is insignificant.
RNA integrity is verified by capillary electrophoresis using an Agilent 2100 Bioanalyzer instrument. FirstChoice total RNA is provided in THE RNA Storage Solution at a convenient, ready-to-use concentration of 1 mg/mL.
Ordering Information
NOTE: Ambion no longer sells the following purified RNAs/cDNAs as catalog made-to-stock products:
- FirstChoice Tumor/Normal Adjacent Tissue RNA
- FirstChoice Brain Region RNA
- FirstChoice Rat/Mouse RNA
- FirstChoice Poly(A) RNA
- FirstChoice cDNA (either PCR-ready or RACE-ready)
To order any of the above products please contact customer service.
Viral RNA From Liquid & Cell-Free Samples
Viral analysis of biological (animal, insect, plant, fungal, bacterial) and environmental (water, air, food) samples is increasingly conducted using molecular testing strategies that require viral RNA or DNA from the samples being tested. Purification of viral RNA poses unique challenges for getting good recovery and detection sensitivity. Ambion RNA experts have developed RNA purification products that are optimized to provide maximum viral RNA yield, purity, and integrity from a broad range of sample types in several format options.
Product Specifications
Product | Format | Most Suitable Downstream Application | ||||
---|---|---|---|---|---|---|
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
Total RNA | ||||||
PureLink Viral RNA⁄DNA Mini Kit | Chaotropic salt-based lysis, silica filter | |||||
RNAqueous Kit | Chaotropic salt-based lysis, silica filter | |||||
MagMAX AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | |||||
MagMAX-96 AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
DNA/RNA | ||||||
Product | Format | Most Suitable Downstream Application | ||||
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
PureLink Pro 96 Viral RNA⁄DNA Purification Kit | Chaotropic salt-based lysis, silica filter, 96-well processing, automation compatible | |||||
MagMAX Total Nucleic Acid Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
MagMAX-96 Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
Dynabeads SILANE Viral NA | Chaotropic salt-based lysis, magnetic beads, automation compatible | |||||
iPrep PureLink Virus Kit | Chaotropic salt-based lysis, magnetic particles, automation compatible | |||||
MagMAX Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | |||||
MagMAX-96 Viral RNA Isolation Kit (5 x 96 rxns) | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
MicroRNA | ||||||
Product | Format | Most Suitable Downstream Application | ||||
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
mirVana miRNA Isolation Kit | Chaotropic salt-based lysis, silica filter | |||||
mirVana miRNA Isolation Kit (without phenol - must be purchased separately) | Chaotropic salt-based lysis, silica filter | |||||
TRIzol LS Reagent | Lysis in TRIzol reagent, one-step reagent | |||||
mRNA | ||||||
Product | Format | Most Suitable Downstream Application | ||||
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
Dynabeads mRNA DIRECT Kit | Chaotropic salt-based lysis, magnetic beads | |||||
Dynabeads mRNA DIRECT Micro Kit | Chaotropic salt-based lysis, magnetic beads |
Total RNA Extraction
Starting with high quality, pure, and intact total RNA is critical to many experiments, (e.g. RT-PCR, qRT-PCR, array analysis, Northern blots, nuclease protection assays and RNA sequencing). The table below will help you choose the right Invitrogen product to purify total RNA from your specific sample type and sample size.
Which Total RNA Purification Kit is right for you?
Order now | Order now | Order now | Order now | |
Gold standard for highly pure, intact RNA | High-quality RNA in less than 20 minutes | High-throughput purification of RNA and DNA | Complete, no purification system for qRT-PCR results | |
---|---|---|---|---|
TRIzol Reagents | PureLink Kits | MagMAX Kits | Cells to Ct | |
Recommendations | Most Cited, Top Seller | Best for qPCR
| ||
Isolation method | High purity organic extraction (requires alcohol precipitation) | Fast, convenient silica column | Scalable, flexible format with magnetic beads | Chemical lysis (time to qRT-PCR results) |
Prep time | ~1 hr | < 20 minutes | ~45 minutes | 10 minutes
|
Compatible sample types | Most sample types, particularly more difficult to lyse | Cells and tissues | Tissues, cells, blood, serum/plasma, urine, plants | Cells
|
Amount of starting material | 100 mg of tissue or 107 cells (requires 1 mL reagent) | Up to 200 mg tissue, 5x106 to 1x108 cells | Up to 100 mg tissue, Up to 5x106 cells | 1–100,000 cells
|
Reverse transcription reagents included | | |||
qPCR Master mix included | ||||
High throughput compatible | (plate format available) |
Organic extraction & purification
Organic solvent extraction offers tried-and-true performance, and is ideal for the most stringent applications when combined with silica-membrane filtration or magnetic bead purification. Invitrogen TRIzol Reagent offers:
- Broadest sample type compatibility
- Robust method
- Good for difficult samples (e.g. high lipid tissue) and scalable sample input
- Wide acceptance as the gold standard
Magnetic bead purification
Magnetic particles offer solution-phase binding and washing kinetics to facilitate RNA purification and improved sample handling, including automation, ideal for higher throughput processing. Applied Biosystems MagMAX Magnetic Beads offer:
- Flexible and scalable throughput
- Amenable to automation for higher throughput
- Better removal of inhibitors and low elution volume resulting in more concentrated RNA
Invitrogen RNA essentials
Invitrogen products, are designed to minimize nuclease contamination and ensure experimental success in less-than-ideal conditions. These products are manufactured under the most stringent quality specifications and include:
- RNA purification kits
- Specimen collection and RNA stabilization solutions
- Nuclease inhibitors and decontaminants
- Nuclease-free buffers and plastics
- RNA ladders
- Northern blot reagents
Silica-membrane purification
Silica-membrane filters allow fast and easy RNA purification resulting in pure RNA that can be used in most applications. Invitrogen PureLink purification columns offer:
- A fast and easy method suitable for most downstream uses
- Wide sample type compatibility
- Familiar spin column format commonly used for low-med throughput
Total Nucleic Acid Extraction / Protein
Research sometimes requires testing of more than just RNA from each sample. For example researchers may want to compare the effects of genetic variations on gene expression, or to evaluate both protein and cognate mRNA levels. To serve this need, some Ambion purification products are designed to extract both RNA and DNA, or both RNA and protein from samples. Kits with these capabilities allow users to get more out of their samples while employing one kit to streamline the process.
The MELT Total Nucleic Acid Isolation Kit provides a method to obtain RNA or DNA from fresh or frozen tissue without physical digestion. For FFPE samples, the RecoverALL Total Nucleic Acid Isolation Kit for FFPE sets the standard and allows users to purify both RNA and DNA via a conventional filter-based method, while the new MagMAX FFPE Total Nucleic Acid Isolation Kit offers the same RNA and DNA purification from FFPE samples in a magnetic bead format without the need for xylene.
Nucleic Acid & Protein Extraction Kits
The PARIS Kit is designed to simultaneously purify total RNA and generate a native lysate suitable for common protein analysis techniques. The related mirVana PARIS Kit can also be used to obtain both total RNA and native lysate, but it also recovers the small RNA fraction, either along with total RNA or as a fraction enriched for small RNAs.
Total Nucleic Acid Extraction Kits
The MagMAX Total Nucleic Acid Isolation Kit purifies both RNA and genomic DNA with a combined mechanical (zirconia-bead beating) and chemical lysis method designed specifically for hard-to-lyse bacteria. Conversely, the MELT Total Nucleic Acid Isolation Kit provides a closed-tube, enzymatic method to obtain RNA or DNA from fresh or frozen tissue without physical disruption. For FFPE samples, the RecoverALL Total Nucleic Acid Isolation Kit for FFPE is designed to purify both RNA and DNA via a conventional glass fiber filter-based method, while the new MagMAX FFPE Total Nucleic Acid Isolation Kit offers xylene-free RNA and DNA purification from FFPE samples using a magnetic bead method.
Publication
MELT Total Nucleic Acid Isolation System: a new technology for hands-free tissue disruption, RNA preservation and total nucleic acid purification. Nature Methods, 23 Aug 2005.
microRNA & small RNA Isolation
MicroRNAs (miRNAs) and small interfering RNAs (siRNAs) modulate gene expression. Along with several other classes of small noncoding RNAs (e.g. snRNA, snoRNA and piRNA), they have tremendous impact on a variety of biological processes.
While most traditional RNA isolation methods are not very efficient at recovering these smaller RNAs, several kits developed by our RNA experts offer quantitative recovery of small RNAs from a variety of sample types.
Which microRNA & small RNA isolation kit is right for you?
Order now | Order now | Order now | Order now | |
30 min isolation from most samples | Recover all RNAs as well as proteins | HTP isolation of total RNA enriched for small RNAs | Complete kit, sample-to-qPCR from cells | |
---|---|---|---|---|
mirVana miRNA Isolation Kit (with or without phenol) | mirVana Paris | MagMAX mirVana Total RNA Isolation Kit | TaqMan microRNA Cells-to-CT Kit | |
Top Seller | ||||
RNA types isolated | Small & large RNA molecules | Small & large RNA molecules | Small & large RNA molecules | Small & large RNA molecules |
Top Seller | ||||
Prep time | 20 minutes | 30 minutes | <1 hour | 10 minutes |
Compatible sample types | Most sample types | Most sample types | Serum/Plasma, whole blood, Fresh/frozen tissue, cells, urine | Cells |
Isolation method | Highest purity and convenience; includes both organic extraction and silica column | Highest purity and convenience; includes both organic extraction and silica column | Scalable, flexible format with magnetic beads | Cells-to-CT direct lysis, no purification required |
Reverse transcription reagents included | No | No | No | TaqMan MicroRNA Reverse Transcription Kit |
qPCR Master mix included | No | No | No | TaqMan |
Kit size | 40 preps | 40 preps | 96 preps | 100 lysis reactions - plus - reagents for 200 qRT-PCR assays |
Publications
- Johnson CD, Esquela-Kerscher A, Stefani G, Byrom M, Kelnar K, Ovcharenko D, Wilson M, Wang X, Shelton J, Shingara J, Chin L, Brown D, Slack FJ. The let-7 MicroRNA Represses Cell Proliferation Pathways in Human Cells. Cancer Res. 2007 Aug 15;67(16):7713-22. [Abstract]
- Shingara J, Keiger K, Shelton J, Laosinchai-Wolf W, Powers P, Conrad R, Brown D, Labourier E. An optimized isolation and labeling platform for accurate microRNA expression profiling.RNA. 2005 Jul 25; [Epub ahead of print] [Abstract]
- Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, Labourier E, Reinert KL, Brown D, Slack FJ. RAS is regulated by the let-7 microRNA family. Cell. 2005 Mar 11;120(5):635-47. [Abstract]
- Cheng AM, Byrom MW, Shelton J, Ford LP. Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res. 2005 Mar 1;33(4):1290-7. [Abstract]
Technical Resources
- Potent Reagents for miRNA Studies
- Getting Started with MicroRNA Research
- MicroRNA Cloning Overview
- Prepare siRNA and miRNA Probes in Just 1 Hour
- Tips from the Bench: The Optimal Thickness of FFPE Sections for miRNA Isolation
- Isolate and Quantitate miRNA from Plants
- Tips from the Bench: High Throughput miRNA Isolation
- Tips from the Bench: Stability of miRNA at Sub-Zero Temperatures
- Your Data: Isolation of miRNAs from Pancreatic Samples
- Isolating miRNA for Profiling Studies
- Extract microRNA from Blood Samples
- Examine microRNA Profiles from Archived, Formalin-fixed, Paraffin-embedded (FFPE) Tissues
- Got Small RNA?
- Get the miRNA Out—Plant Applications
- Definition and Overview
- miRNA Function
- Expression & Targets
- miRNA Processing
mRNA Extraction
Which mRNA purification kit is right for you?
Order now | Order now | Order now | Order now | |
Rapid mRNA purified from total RNA | No purification needed, mRNA direct from crude samples | mRNA from micro-sized samples Best for next-generation sequencing of mRNA | HTP mRNA directly or enrich from total RNA | |
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Dynabeads\ mRNA Purification Kit | Dynabeads mRNA DIRECT | Dynabeads mRNA DIRECT Micro Kit | mRNA Catcher Plus | |
Top seller | ||||
Prep time | 15 minutes | 15 minutes | 15 minutes | 1.5 hours |
Direct isolation of mRNA from crude samples | No | Yes | Yes | Yes |
mRNA enrichment from total RNA | Yes | No | No | Yes |
Amount of starting material | <75 μg Total RNA | <20 x 106 cells 2–200 mg animal tissue 4–400 mg plant tissue | <1 x 104 cells <5 mg plant or animal tissue | 100 to 1 x 106 cells <4 mg tissue 40 µL whole blood 100 ng–100 µg total RNA |
Format | Fast magnetic bead capture from pure total RNA | Fast magnetic bead capture directly from crude samples | Fast magnetic bead capture directly from micro-size samples | HTP ready 96-well coated plate |
High-throughput compatible | Yes | Yes | Yes | Yes |
Kit size | 2 mL oligo dT beads | 5 mL or 10 mL oligo dT beads | 2 mL oligo dT beads (sufficient for 100 mRNA preps) | 96 or 960 preps |
Magnetic Bead mRNA Extraction and Enrichment Kits
Oligo(dT)/Magnetic Capture Methods
Column methods for mRNA purification are reliable but require extensive manual interaction, which means more time and reduced throughput. Magnetic beads conjugated with oligo(dT) offer additional handling and scaling benefits; however, this procedure does require the use of magnetic stands.
The Invitrogen Ambion Poly(A)Purist MAG Kit employs the same hybridization and wash solutions as those in the Poly(A)Purist Kit but with the added ease, speed, and scalability of magnetic beads. The protocol typically takes only 45 minutes, and the kit performs from 8 large enrichment reactions (up to 1 mg total RNA input) to 80 small enrichment reactions (as little as 30 µg total RNA input). Like the Poly(A)Purist Kit, this kit only enriches mRNA from previously purified total RNA.
The Invitrogen Dynabeads mRNA Purification Kit for mRNA Purification from Total RNA Preps, enriches mRNA from purified total RNA, typically in 15 minutes, and allows users the flexibility to elute the mRNA from the beads in as little as 5 µL, or even to perform cDNA synthesis directly on the beads without elution.
For those who prefer using Dynabeads technology with sample lysis and enrichment in one kit, the Invitrogen Dynabeads mRNA DIRECT Kits enable 15-minute lysis and enrichment processing of a variety of sample types. In fact, it is possible to capture mRNA and generate cDNA libraries from a single cell using this technology. The flexible format allows users to scale up or down and gives the option to elute the mRNA from the beads in as little as 5 µL, or to perform cDNA synthesis directly on the beads without elution. The 5 mL size is sufficient for 20 standard isolations, while the 10 mL size is sufficient for 40 standard isolations.
For those who work with smaller inputs, the Invitrogen Dynabeads mRNA DIRECT Micro Kit provides enough reagents for 100 mRNA isolations from up to 2.5 x 104 mononuclear cells, up to 1 x 104 cultured cells, or up to 5 mg tissue (depending on the tissue), with the same elution flexibility as the other Dynabeads kits.
Bacteria lack the relatively stable poly(A) tails found on eukaryotic mRNA. Until very recently, isolating mRNA from bacteria has been virtually impossible. The Invitrogen MICROBExpress Bacterial mRNA Isolation Kit employs a novel technology to remove up to >95% of the 16S and 23S rRNA from total RNA of E. coli and other bacterial species. The kit is suitable for mRNA enrichment from ≤10 µg of purified bacterial total RNA in about 2 hours from a broad spectrum of gram-positive and gram-negative bacteria. mRNA isolated with the MICROBExpress Kit is a superior template for synthesizing labeled cDNA for array analysis and is ideal for quantitative RT-PCR, northern blotting, and cDNA library construction.
mRNA Extraction and Enrichment via Coated Plates
The most distinctive mRNA purification method takes advantage of specially treated plates in conjunction with optimized reagents to both lyse and enrich mRNA from cells, tissue, blood, or purified total RNA. The Invitrogen mRNA Catcher PLUS product utilizes a proprietary surface treatment of a 96-well plate to facilitate easy and automation-friendly mRNA purification. Locked Nucleic Acid (LNA) technology incorporated into the 20-mer oligo(dT) increases specificity and hybridization efficiency while removing centrifugation, precipitation, and phase-separation steps from the overall workflow.
The mRNA Catcher PLUS Kit is capable of handling 100 ng to 100 µg total RNA, 100 to 106 mammalian cells, up to 4 mg tissue, and 40 µL whole blood. The binding capacity is >80 ng/well. The protocol employs a simple lysis, hybridization, wash, and elution process that can be completely automated on open-format liquid handlers in approximately 1.5 hours from start to finish. The system also has the flexibility to elute mRNA or to perform cDNA synthesis directly in the hybridization plate without elution.
Sequence-Specific RNA Extraction
Target a specific RNA sequence and isolate only those RNA molecules directly from crude lysates or other biological fluids using Dynabeads. Captured RNA is suitable for a wide range of downstream applications, including Dynabeads Streptavidin are ideal for numerous applications, including purification of proteins, nucleic acids purification, protein interaction studies, immunoprecipitation, immunoassays, phage display, biopanning, drug screening and cell isolation.
Which Sequence-Specific RNA Purification Kit or Reagent is Right for You?
Order now | Order now | Order now | |
4 beads in one kit, identify best for your application | Best solution for viscous samples | High surface area for maximum enrichment | |
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Dynabeads Streptavidin Trial Kit | Dynabeads M-270 Streptavidin | Dynabeads MyOne Streptavidin C1 | |
Top seller | |||
Bead size | 1 µm x 2, 2.8 µm x 2 | 2.8 µm | 1 µm |
Biotinylated Ig (µg/mg beads) | up to 20 | up to 10 | up to 20 |
Binding Capacity (Free Biotin) | 6502,500 pmoles/mg beads | 650–1,350 pmoles/mg beads | 2,500 pmoles/mg beads |
Pack size | 4 x 1 mL | 2 or 10 mL | 2 or 10 mL |
Ligand type | Streptavidin | Streptavidin | Streptavidin |
Dynabeads Technology
Our Streptavidin-coupled Dynabeads are a robust and versatile tool that can be used to literally target and capture specific RNA or DNA sequences and then pull them directly out of solution. These monosized superparamagnetic Dynabeads provide an efficient and solid-phase alternative to nitrocellulose and provide you with an unmatched level of product quality and data consistency. Excellent near liquid phase reaction kinetics allow for extremely fast protocols. The inherent ease of magnetic handling mean that downstream manipulations and buffer changes are as simple as concentrating the bead-bound target at the tube-wall with a magnet and then discarding the supernatant. These beads are compatible with an extremely broad range of sample types including most bodily fluids, crude lysates of plant, animal and microbial origin as well as purified total RNA or DNA. Since these Dynabeads will only interact with specifically targeted RNA or DNA molecules, upstream purification of total RNA or DNA is almost always an unnecessary step.
The 1 µm Dynabeads MyOne Streptavidin C1 present a very high surface area per mg of beads, enabling high enrichment of low abundance RNA or DNA. When the goal is to capture nucleic acid from more viscous samples such as cerebrospinal fluid, the larger 2.8 µm sized Dynabeads M-270 Streptavidin are recommended. These Dynabeads (MyOne Streptavidin C1 and M-270 Streptavidin) are optimally designed to have slightly negatively charged surfaces which ensure negligible non-specific binding of non-target nucleic acid sequences.
Examples of applications include; isolation of RNA/DNA infectious agent (1,2,3,4), subtractive hybridization (5,6,7), cDNA selection and enrichment, detection and isolation of mutated sequences (8,9,10), isolation of cell specific transcripts and mRNA differential display.
Learn more about:
The direct capture procedure involves the immobilization of double-stranded PCR products onto the beads. These are easily converted to single-stranded bead-bound templates which are then used to capture specific RNA or DNA molecules directly from solution.
An alternative indirect capture approach will offer faster reaction kinetics in some cases. This indirect capture procedure allows the target sequence to be captured prior to being immobilized the magnetic beads. First, a biotinylated capture-sequence (single-stranded DNA) is incubated with the sample and allowed to hybridize to the targeted RNA or DNA molecules in solution. Streptavidin coated Dynabeads are then added to the mixture and the hybridized sequences are immobilized onto the Dynabeads via the streptavidin-biotin bond.
Did you know?
Did you know that Dynabeads are employed on more than 25,000 routine IVD instruments worldwide?
Selected References
- Meng Q. et al. (2001) Automated multiplex assay system for simultaneous detection of hepatitis B virus DNA, hepatitis C virus RNA and human immunodeficiency virus type 1 RNA. J.Clin.Microbiol. 39(8):2937-2945.
- Stevens SJC. et al. (1999) Monitoring of Epstein-Barr virus DNA load in peripheral blood by quantitative competitive PCR. J.Clin. Microbiol. 37:2852-2857.
- Mangiapan G. et al. (1996) Sequence capture-PCR improves detection of mycobacterial DNA in clinical specimens. J. Clin. Microbiol. 34(5):1209-1215.
- Shuber AP. et.al. (2002) Accurate, noninvasive detection of Helicobacter pylori DNA from stool samples: potential usefulness for monitoring treatment. J. Clin. Microbiol. 40(1):262-264.
- Hansen-Hagge TE. et.al. (2001) Identification of sample-specific sequences in mammalian cDNA and genomic DNA by the novel ligation-mediated subtraction (LIMES). Nucl. Acids Res. 29(4):e20.
- Pradel N. et.al. (2002) Genomic subtraction to identify and characterize sequences of Shiga toxin-producing Escherichia coli O91:H21. Appl. Env. Microbiol. 68(5):2316-2325.
- Laveder P. et.al. (2002) A two-step strategy for constructing specifically self-subtracted cDNA libraries. Nucleic Acids Res. 30(9):e38.
- Lindblad-Toh K.et.al. (2000) Large-scale discovery and genotyping of single-nucleotide polymorphisms in the mouse. Nature Genetics. 24:381-386.
- Miyashiro I. et.al. (2001) Molecular strategy for detecting metastatic cancers with use of multiple tumor-specific MAGE-A genes. Clin.Chem. 47(3):505-512.
- Dong SM. et.al. (2001) Detection of colorectal cancer in stool with the use of multiple genetic targets. J Natl. Cancer Inst. 93(11):858-865.
Purified RNA
Ambion RNA experts are committed to providing the highest-quality prepared RNA available. To achieve this goal we are continually striving to understand what "high-quality RNA" truly means with respect to various downstream applications. Intactness of mRNA, genomic DNA contamination, presence/absence of enzymatic inhibitors, and ribosomal RNA contamination in poly(A) preparations are all important aspects of RNA quality. As we continue to gain a better understanding of how these factors affect downstream applications, we will strive to ensure that our FirstChoice RNA products exceed the requirements of even the most stringent applications.
- Certified to contain small RNAs (miRNA, siRNA, and snRNA)
- RNA available from a wide variety of human tissues and cell lines
- DNase-treated to remove contaminating DNA
- Superior quality control standards
Total RNA
We offer FirstChoice total RNA from human, mouse, and rat tissues, as well as from E. coli and human cell lines. RNA is obtained using Ambion RNA isolation reagents to yield highly pure, intact RNA that includes the small-RNA fraction. Purified RNA is subjected to a stringent DNase treatment. The RNA is supplied ready to use in any downstream application, including RT-PCR. Note that although it is impossible to remove every DNA molecule from an RNA sample, each preparation is tested to ensure that any residual DNA contamination is insignificant.
RNA integrity is verified by capillary electrophoresis using an Agilent 2100 Bioanalyzer instrument. FirstChoice total RNA is provided in THE RNA Storage Solution at a convenient, ready-to-use concentration of 1 mg/mL.
Ordering Information
NOTE: Ambion no longer sells the following purified RNAs/cDNAs as catalog made-to-stock products:
- FirstChoice Tumor/Normal Adjacent Tissue RNA
- FirstChoice Brain Region RNA
- FirstChoice Rat/Mouse RNA
- FirstChoice Poly(A) RNA
- FirstChoice cDNA (either PCR-ready or RACE-ready)
To order any of the above products please contact customer service.
Viral RNA From Liquid & Cell-Free Samples
Viral analysis of biological (animal, insect, plant, fungal, bacterial) and environmental (water, air, food) samples is increasingly conducted using molecular testing strategies that require viral RNA or DNA from the samples being tested. Purification of viral RNA poses unique challenges for getting good recovery and detection sensitivity. Ambion RNA experts have developed RNA purification products that are optimized to provide maximum viral RNA yield, purity, and integrity from a broad range of sample types in several format options.
Product Specifications
Product | Format | Most Suitable Downstream Application | ||||
---|---|---|---|---|---|---|
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
Total RNA | ||||||
PureLink Viral RNA⁄DNA Mini Kit | Chaotropic salt-based lysis, silica filter | |||||
RNAqueous Kit | Chaotropic salt-based lysis, silica filter | |||||
MagMAX AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | |||||
MagMAX-96 AI⁄ND Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
DNA/RNA | ||||||
Product | Format | Most Suitable Downstream Application | ||||
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
PureLink Pro 96 Viral RNA⁄DNA Purification Kit | Chaotropic salt-based lysis, silica filter, 96-well processing, automation compatible | |||||
MagMAX Total Nucleic Acid Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
MagMAX-96 Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
Dynabeads SILANE Viral NA | Chaotropic salt-based lysis, magnetic beads, automation compatible | |||||
iPrep PureLink Virus Kit | Chaotropic salt-based lysis, magnetic particles, automation compatible | |||||
MagMAX Viral RNA Isolation Kit | Chaotropic salt-based lysis, magnetic particles | |||||
MagMAX-96 Viral RNA Isolation Kit (5 x 96 rxns) | Chaotropic salt-based lysis, magnetic particles, 96-well processing, automation compatible | |||||
MicroRNA | ||||||
Product | Format | Most Suitable Downstream Application | ||||
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
mirVana miRNA Isolation Kit | Chaotropic salt-based lysis, silica filter | |||||
mirVana miRNA Isolation Kit (without phenol - must be purchased separately) | Chaotropic salt-based lysis, silica filter | |||||
TRIzol LS Reagent | Lysis in TRIzol reagent, one-step reagent | |||||
mRNA | ||||||
Product | Format | Most Suitable Downstream Application | ||||
Real-Time RT-PCR | Microarray Analysis | Sequen-cing | Northern Blot | Cloning | ||
Dynabeads mRNA DIRECT Kit | Chaotropic salt-based lysis, magnetic beads | |||||
Dynabeads mRNA DIRECT Micro Kit | Chaotropic salt-based lysis, magnetic beads |
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For Research Use Only. Not for use in diagnostic procedures.