Shop All RNA Purification Kits

LeukoLOCK™ Total RNA Isolation System (Invitrogen™)

The LeukoLOCK™ Total RNA Isolation System is an innovative method for cellular fractionation of whole blood and total RNA stabilization and extraction from the leukocyte population. The kit is comprised of a LeukoLOCK™ Fractionation & Stabilization Kit (also available separately; Cat. No. AM1933) and a LeukoLOCK™ Total RNA Isolation Kit.

• Fractionates leukocytes from whole blood in minutes
• Extracts total RNA from leukocytes
• No additional globin reduction steps required
• Stabilizes RNA to preserve expression profile
• Preserves leukocytes at room temperature for days

The LeukoLOCK™ system is optimized for use with human blood. Blood is a storehouse of cellular information; however, the presence of globin mRNA in RNA prepared from whole blood can interfere with downstream expression profiling applications. The LeukoLOCK™ system employs filter-based leukocyte-depletion technology to isolate leukocytes from whole blood and RNAlater® solution to stabilize the cells on the filter. By excluding red blood cells, the RNA that is purified from captured leukocytes is inherently depleted of globin mRNA, which improves sample utility for expression profiling and other applications (see Figure).

Leukocyte Capture and Stabilization
A 9–10 mL sample of anticoagulated blood is passed through a LeukoLOCK™ filter, which captures the total leukocyte population while eliminating red blood cells (including reticulocytes), platelets, and plasma. After rinsing with phosphate-buffered saline, the filter is flushed with RNAlater® solution to stabilize the RNA in the captured leukocytes (see protocol schematic). The RNA can be isolated immediately, or stabilized cells can be maintained for several days at room temperature or for longer periods at –20°C or –80°C (see Figure).

RNA Isolation
Purification of the RNA is done by first disrupting the captured cells in a guanidinium thiocyanate–based solution that releases the RNA while simultaneously inactivating nucleases. The cell lysate is collected and briefly treated with Proteinase K. RNA is then purified from the lysate using MagMAX™ magnetic bead-based technology for washing, followed by DNase treatment (with TURBO DNase™) and final clean-up.

Expression Studies
Both quantitative RT-PCR and microarray expression studies have validated the suitability of the purified RNA for reliable transcriptome profiling (see Figure). Compared to RNA processed with a commonly used blood RNA isolation procedure, RNA isolated with the LeukoLOCK™ system produces longer median aRNA after amplification and higher percent present calls without requiring additional post-extraction steps to remove globin mRNA (see Figure). Since globin aRNA represents 25–40% of the aRNA peak from whole blood total RNA, the yield of total RNA is understandably lower with methods that deplete unwanted globin transcripts. RNA amplified after LeukoLOCK™ purification offers greater array sensitivity due to the dramatic reduction in competing globin transcripts.

Kit Yields
Recovery varies from donor to donor, but is typically 10–20 µg of RNA per 9–10 mL of whole blood (see Figure). RNA recovered using the LeukoLOCK™ system contains less than 1/10th the amount of reticulocyte-derived alpha- and beta-globin mRNAs present in typical RNA samples from unfractionated whole blood.

Dynabeads™ mRNA DIRECT™ Purification Kit (Invitrogen™)

The Dynabeads® mRNA DIRECT™ Kit is designed for simple and rapid isolation of pure, intact polyadenylated (polyA) mRNA directly from the crude lysate of animal and plant cells and tissues. The isolated mRNA is suitable for use in all downstream applications. Advantages of the Dynabeads® mRNA DIRECT™ Kit:

Fast—15 minute procedure yields pure, intact mRNA
Highly pure mRNA isolation—best choice upstream of cDNA synthesis
Sensitive mRNA isolation—enables cDNA synthesis and cDNA library construction from ultra-small starting samples (enables cDNA library construction from a single cell)

System overview
The isolation protocol relies on base pairing between the polyA residues at the 3' end of most mRNA, and the oligo (dT)25 residues covalently coupled to the surface of the Dynabeads® . Other RNA species lacking a polyA tail will not hybridize to the beads and are readily washed away. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. RNase inhibiting agents in the Lysis/Binding Buffer together with stringent hybridization and washing conditions ensure the isolation of pure, intact mRNA from crude samples rich in RNase, without the use of strong chaotropic agents. The mRNA purification beads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples (see protocol). 1 mg of Dynabeads® Oligo (dT)25 beads (200 µL) binds up to 2 µg of mRNA. A typical mammalian cell contains about 10–30 pg of total RNA, from which 1–5% is mRNA.

RiboPure™ RNA Purification Kit (Invitrogen™)

The Ambion® RiboPure™ Kit is designed for rapid purification of high-quality RNA from tissue samples or cultured cells. The kit combines TRI Reagent® with glass fiber filter purification to yield exceptionally pure RNA, free of residual proteins and lipids. The kit includes sufficient reagents for 50 purifications from 1–100 mg of tissue or 0.1–20 x 106 cultured cells each.

• Yields the highest purity RNA attainable
• Ideal for the most stringent downstream applications
• Handles problematic samples with ease
• Fast (less than 30 minutes) and easy procedure

RNA isolated using the RiboPure™ Kit is suitable for use in all common downstream applications, including cDNA synthesis, real-time and end-point RT-PCR, microarray analysis, northern blots, and RNase protection assays. The RiboPure™ protocol is especially suited for processing samples that are difficult to disrupt, or samples that have large amounts of ribonuclease or other contaminants. Samples are first homogenized in TRI Reagent®, a monophasic solution containing phenol and guanidine thiocyanate, to rapidly lyse cells and inactivate nucleases. Addition of bromochloropropane (BCP) results in separation of the homogenate into aqueous and organic phases. RNA partitions to the aqueous phase, while DNA and protein remain in the interphase and organic phase. The aqueous phase is removed and passed through a glass fiber filter that binds the RNA. The filter is washed, and the RNA is eluted. The procedure is compatible with tissues that have been stored in Ambion's RNAlater®. Total RNA yield is typically 100–500 µg per 100 mg of tissue, depending on the type of tissue.

mRNA Catcher™ PLUS Purification Kit (Invitrogen™)

The mRNA Catcher™ PLUS Kit enables high-throughput purification of mRNA from cells, tissues, blood, and total RNA in a 96-well format. By taking advantage of Locked Nucleic Acid (LNA) technology, the mRNA Catcher™ PLUS Kit provides improved performance and a faster protocol, while preserving mRNA yield and purity (see table). The mRNA Catcher™ PLUS Kit offers:

Robust performance—high binding capacity per well (>80 ng/well)
Reproducible results—low CVs from well-to-well and plate-to-plate (~2%)
Optimized protocols—for small sample sizes (100 cells, 4 mg tissue, and 40 µL blood)
Compatible plate design—may be used with most PCR cyclers

How it works
mRNA from total RNA, cells, tissue, and blood samples is hybridized to an immobilized, single-stranded 20-mer oligo(dT)-containing LNA. The incorporation of LNA into the oligo probe increases the hybridization efficiency and specificity of the mRNA binding. The mRNA can either be eluted or directly reversed-transcribed to cDNA directly in the well. The mRNA Catcher™ PLUS method does not require centrifugation, precipitation, phase separation, or column filtration. In addition, the entire method is amenable to automation for streamlined, high-throughput expression profiling for large-scale processes. Isolated mRNA is suitable for use in downstream applications such as RT-PCR and qRT-PCR.

PureLink™ FFPE RNA Isolation Kit (Invitrogen™)

The PureLink® FFPE RNA Isolation Kit provides a simple, reliable, and rapid method to isolate high-quality total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue specimens for retrospective basic and clinical biological studies of gene expression without the use of toxic organic solvents for deparaffinization.
• Offers a rapid and environmentally friendly procedure for the extraction of RNA from FFPE tissue specimens
• Isolates RNA with maximum yield and minimum degradation

Rapid and Non-Toxic Isolation Procedure
The PureLink® FFPE RNA Isolation Kit employs a simple heating procedure for deparaffinization, instead of relying on the use of toxic organic solvents such as xylene. RNA isolation can be completed in 30 - 40 minutes instead of days, as is required for older FFPE isolation methods (Figure 1). The heating procedure for deparaffinization partly restores RNA template structure for cDNA synthesis and improves template quality for reverse transcription.

High Yield, High Quality RNA
The PureLink® FFPE RNA Isolation Kit offers higher yields and purity as compared to other commercially available RNA purification systems, allowing for RT-PCR products over 1 kb in length (Figure 2). In addition, the PureLink® FFPE RNA Isolation Kit minimizes genomic DNA contamination of the purified RNA sample and provides an option for DNase digestion.

Purified Total RNA is Suitable for Use with a Variety of Downstream Applications and Products
RNA extracted from FFPE specimens is suitable for downstream applications such as reverse transcription, real-time PCR (qPCR), real-time RT-PCR (qRT-PCR), and microarray analysis.

For Research Use Only. Not intended for animal or human diagnostic or therapeutic use.

Lysis/Binding Buffer for Dynabeads™ mRNA Purification Kits (Invitrogen™)

This Lysis/Binding Buffer is included in the following Dynabeads™ mRNA purification kits: Cat. Nos. 61011, 61012, and 61021. It is made available separately for applications that require more Lysis/Binding Buffer than is provided in the kit.

RiboPure™ RNA Purification Kit, bacteria (Invitrogen™)

The Ambion® RiboPure™-Bacteria Kit provides a rapid, robust, column-based method for isolating high quality RNA from bacteria. Total RNA can be isolated from the toughest Gram positive bacteria samples thanks to the kit's comprehensive protocol that uses zirconia beads to thoroughly disrupt bacterial cell walls. The RiboPure™-Bacteria Kit protocol also eliminates the need for harsh treatments or enzymatic digestion which may alter gene expression profile.
• Obtain purified bacterial RNA suitable for nearly any downstream application
• Prepare pure RNA samples from the most difficult-to-lyse Gram positive bacteria
• Eliminate enzymatic pretreatments that might otherwise alter gene expression
• Remove contaminating gDNA with included DNA-free™ reagents

A Comprehensive and Convenient Column Purification Procedure
The RiboPure™-Bacteria Kit purifies total RNA from a wide range of bacterial species and has been extensively tested with E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, Campylobacter fetus, and Rhodobacter sphaeroides.

A Streamlined and Effective Method for Bacterial RNA Extraction
The kit protocol begins with thorough cell wall disruption using of zirconia beads, followed by organic extraction of the lysate, and RNA purification on a convenient silica column. In approximately 1 hour, RNA is of superb quality can be isolated from fresh, snap-frozen, or RNAlater® solution-treated bacterial cells (Figures 1 and 2). To help ensure complete removal of genomic DNA, an optional DNA removal step requiring approximately 30 minutes of additional time, can be incorporated. When the RiboPure™-Bacteria kit is used upstream from the MICROBExpress™ Bacterial mRNA Enrichment Kit, the sensitivity of RNA-Seq experiments or array analyses can be dramatically increased (Figure 3).

Purified RNA is Suitable for Use with a Variety of Downstream Applications and Products:
The RNA purified using the Ambion® RiboPure™-Bacteria Kit is suitable for downstream applications including RNA sequencing, microarray analysis, real-time PCR (qPCR), reverse transcription qPCR (RT-qPCR), northern blotting, and cDNA library construction.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

GeneJET Plant RNA Purification Kit (Thermo Scientific™)

Thermo Scientific GeneJET Plant RNA Purification Mini Kit is designed for rapid and efficient purification of high quality total RNA from wide variety of plant species and tissue types.

The kit utilizes silica-based membrane technology in the form of a convenient spin column, eliminating the need for expensive resins, toxic phenol-chloroform extractions, or time-consuming alcohol precipitation. The standard procedure takes less than 20 minutes following cell lysis. Purified high quality RNA can be used in a wide range of downstream applications, such as RT-PCR, RT-qPCR, Northern blotting and other RNA-based analysis. RNA yields vary between different species, tissues, and age of tissue sample. See Table 1 for typical total RNA yields from various sources.

Highlights

Fast—procedure completed in 20 min following cell lysis
Convinient—protocol does not include clogging-prone lysate filtration step
High yield—up to 65 µg of high purity and high integrity RNA from 50 mg plant sample
Compatible with wide variety of samples (leaves, roots, sprouts and other parts of various plants)

Applications

• Fast extraction of high purity RNA suitable for all conventional molecular biology procedures, including:
• RT-PCR
• RT-qPCR
• Northern blotting
• Nuclease protection assay

GLOBINclear™ Kit, human, for globin mRNA depletion (Invitrogen™)

The GLOBINclear™-Human Kit uses a novel, non-enzymatic technology that rapidly depletes >95% of the alpha and beta globin mRNA from total RNA preparations derived from whole blood. Advantages of the GLOBINclear™-Human Kit:

• Removes >95% of unwanted globin mRNA from whole blood total RNA
• Detects up to 50% more genes—analyze previously undetected genes
• Achieves greater representation of the genome, low 3'/5' ratios
• Avoids RNase H treatments that can degrade mRNA and alter expression profiles

Using whole blood total RNA samples
It is widely known that expression array data generated from whole blood total RNA samples have reduced detection sensitivity compared to data from fractionated blood samples. Examination of whole blood total RNA reveals a dominant band in the 600–700 base region. This band represents globin mRNA, which is expressed at high levels in red blood cells (RBCs) and reticulocytes (RBC precursors). Up to 70% of the mRNA (by mass) in whole blood total RNA are globin transcripts. Array analysis using total RNA containing a high percentage of globin mRNA has been shown to decrease present calls, decrease call concordance, and increase signal variation.

Using the GLOBINclear Kit
The GLOBINclear™ kit uses a novel hybridization technology that takes advantage of the strength of biotin/streptavidin binding, the specificity of nucleic acid hybridization, and the convenience of magnetic separations. There are no harsh RNase H enzymatic treatments that may compromise the expression profile by degrading mRNAs other than globin mRNA. The resulting mRNA is a superior template for synthesizing labeled cDNA for array analysis and is ideal for quantitative RT-PCR. The enrichment delivers a significant increase in Percent Present calls, decreased sample-to-sample variability, and low 3'/5' ratios.

Accessory product
The kit requires the use of a magnetic stand. The DynaMag-2 Magnetic Stand (Cat. No. 12321D) is recommended.

RNAqueous™-96 Automated Kit (Invitrogen™)

The Ambion® RNAqueous®-96 Automated Kit is used for automated, phenol-free total RNA isolation from 100-500,000 cells or 0.1–1 mg tissue using a guanidinium-based lysis/denaturant and glass fiber filter separation technology. The kit contains enough reagents for 384 (4 x 96) RNA purifications. The RNAqueous®-96 Automated Kit uses an RNA-binding glass-fiber filter to provide high, reproducible yields of intact RNA. An optional on-the-filter DNase treatment can be performed to ensure removal of genomic DNA for RT-PCR applications. Downloadable protocols for the use of this kit with specific liquid handling systems are available at our Automation Resource.

Cells-to-CT™ Bulk Fast Advanced RT Reagents (Invitrogen™)

Invitrogen Fast Advanced Cells-to-CT Bulk RT Reagents are separately-available, large-quantity versions of the reagents used for reverse transcription (RT) in the TaqMan Fast Advanced Cells-to-CT Kit and the SYBR Green Fast Advanced Cells-to-CT kit (20X RT enzyme mix and 20X RT buffer). These reagents are sufficient for 2500 RT reactions. When combined with the Cells-to- CT Bulk Lysis Reagents, these reagents can be easily incorporated into automated, high-throughput applications.

Wash Buffer A for Dynabeads™ mRNA Purification Kits (Invitrogen™)

This Wash Buffer A is included in the following Dynabeads™ mRNA purification kits: Cat. Nos. 61011, 61012, and 61021. It is made available separately for applications that require more Wash Buffer A than is provided in the kit.

Dynabeads™ mRNA DIRECT™ Micro Purification Kit (Invitrogen™)

The Dynabeads® mRNA DIRECT™ Micro Kit is designed for simple and rapid isolation of pure, intact polyadenylated (poly(A)) mRNA. The kit isolates highly purified and intact mRNA directly from cells, tissue, and total RNA samples.

• Obtain extremely pure mRNA in about 15 minutes
• Integrate total RNA purification and mRNA enrichment steps
• Perform reverse transcription and PCR amplification right on the bead
• Use in whole transcriptome library preparation with the Ion Total RNA-Seq Kit v2.0

Rapid Isolation of High Purity mRNA Directly from Samples
The Dynabeads® mRNA Direct Kit contains magnetic beads for the isolation the mRNA transcriptome from a wide variety of samples. Isolation of intact mRNA is possible thanks to RNase inhibitiors in the Lysis/Binding buffer, combined with stringent hybridization and washing steps. Ribosomal RNA and small RNA molecules (transfer RNA, microRNA, small nucleolar RNA, and small cytoplasmic RNA) do not bind to the beads and are eliminated from the preparation. Only polyadenylated RNA species (mRNA) are captured, resulting in cleaner preparations and more sensitive results. Within minutes, pure mRNA is isolated and ready for use in downstream applications.

A Straightforward Enrichment Procedure
The Dynabeads® mRNA DIRECT™ Micro Kit uses a robust affinity purification principle for the enrichment of polyadenylated mRNA. Superparamagnetic Dynabeads®, coupled to oligo-(dT)25, are first equilibrated with Lysis/Binding Buffer, and then mixed with cell or tissue lysate in order to bind mRNA. The beads are then washed to remove contaminating RNA species, and then mRNA is eluted in as little as 5 µl of 10 mM Tris-HCl. The entire process is facilitated by the use of a neodymium magnet (purchased separately), which allows for the quick and efficient immobilization of the magnetic beads during buffer changes.

Designed for a Broad Range of Sample Types
Nearly any biological sample of vertebrate, invertebrate, plant, fungal, or bacterial origin can be used with the Dynabeads® mRNA DIRECT™ Kit. The kit is also ideal for extracting mRNA and viral polyadenylated RNA from blood, serum and sputum. Nucleic acid preparations, including total RNA and unfractionated nucleic acid samples, as well as formalin-fixed, paraffin-embedded (FFPE) tissues, may also be extracted using the Dynabeads® mRNA DIRECT™ Kit.

RNA is suitable for All Downstream Molecular Applications, Including:
• RNA-sequencing
• gene cloning
• cDNA synthesis, cDNA library construction
• RT-PCR, Quantitative RT-PCR
• RPA - Ribonuclease Protection Assay
• Subtractive Hybridization
• Dot/slot Hybridization
• Primer extension

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

RiboPure™ RNA Purification Kit, yeast (Invitrogen™)

For the isolation of extremely high-quality RNA from yeast without the use of enzymatic pretreatments that may alter gene expression profiles. Each Ambion® kit includes sufficient reagents for 50 purifications each from 107 to 108 yeast cells.

• Highest yields of total RNA available
• No enzymatic pretreatment required
• Includes DNA-free™ Reagents

It has traditionally been difficult to isolate high-quality RNA from yeast because of problems disrupting the cells. Many protocols use enzymatic pretreatments that may affect gene expression patterns. The RiboPure™-Yeast RNA Isolation Kit uses no enzymatic pretreatments and thus does not alter expression profiles. It combines a simple and efficient glass bead/one step phenol–mediated disruption using Ambion's Vortex Bead Beater Adapter (see Accessory Products) followed by a glass fiber filter– based RNA purification step. After DNase treatment with the included DNA-free™ Reagents, the resulting RNA is exceptionally pure containing little to no genomic DNA (less than 0.05%). The kit has been used extensively at Ambion to isolate high-quality RNA from Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pichia pastoris, and Candida albicans (see Figure). The kit provides the highest yield of any method tested from each of these species (see Figure).

Accessory Products:
The Kit requires the use of the Vortex Adapter (SKU #AM10024), which turns a standard laboratory vortex into a bead beater. This adapter fits the Vortex Genie® 2 and holds 12 tubes.

MEGAclear™ Transcription Clean-Up Kit (Invitrogen™)

The MEGAclear™ Kit is designed for the quick cleanup of any large-scale transcription reaction. The MEGAclear™ Kit efficiently separates RNA from unincorporated NTPs, enzymes, and buffer components. Advantages of the MEGAclear™ Kit:

• Purifies transcription reactions to produce high-quality RNA, free of unincorporated NTPs, enzymes, and buffer components
• High recovery from 1 ng to 500 µg of input
• Ideal for purification of aRNA amplification reactions, nonisotopically labeled RNA probes, and capped RNA transcripts for oocyte injections
• Quick protocol—purify RNA in less than 30 minutes

Using the MEGAclear™ Kit
The protocol is quick and easy. The transcription reaction is mixed with Binding Solution, passed over an RNA-binding filter, and washed; an elution step releases the purified RNA, which is now ready for any application requiring highly pure synthetic or in vitrotranscribed RNA. For RNA recovery of 70% or greater, single-stranded RNA should be >100 nt and double-stranded RNA should be >200 bp.