Shop All RNA Preparation and Purification

PureLink™ RNA Mini Kit Invitrogen™

The PureLink RNA Mini Kit is a column-based kit used to isolate high-quality total RNA from a wide variety of sample types in 20 minutes using standard laboratory equipment. The kit includes RNase-free lysis and wash solutions that protect RNA from RNases while liberating the RNA from DNA, proteins, and other cellular debris. The fast spin-column workflow is ideal for processing low to mid-throughput batch sizes. The advanced PureLink RNA Mini spin column design allows for maximum sample input (200 mg of tissue) and RNA recovery (up to 1000 µg). This means that users can process both small and large sample sizes with the same RNA isolation kit.

• Process sample inputs from 5–200 mg of animal/plant tissue or from 0.5 x 106 to 108 tissue culture cells with one spin column in 20 minutes
• Isolate more RNA than competitor mini spin columns: up to 1000 µg of total RNA from one column
• Use isolated RNA in most applications without the need for further DNase treatment (very low minimal residual DNA carry over)
• Flexible–optional on-column or post-isolation DNase treatment is available with PureLink DNase Set for easy removal of residual DNA

Extra-large binding capacity enables rapid RNA purification using standard laboratory equipment
The PureLink RNA Mini Kit provides rapid purification of total RNA from a wide range of cells and tissue types to yield up to 1000 µg of purified RNA from a single extraction (see Figure 1). High-quality total RNA can be obtained from mini- to midi-prep amounts of starting material with just trace amounts of residual genomic DNA contamination. The extra-large binding capacity enables one kit to handle most RNA isolation needs in a quick 20 minute protocol without the need for special sample processing instruments.

Easy, optional, on-column DNase treatment for sensitive applications
The PureLink RNA Mini Kit columns are highly efficient for isolating high quality total RNA while removing the majority of genomic DNA. In general, most applications requiring RNA from animal tissue or mammalian cell lines do not require additional DNase treatment. However, some applications such as gene expression analysis by qRT-PCR without intron-spanning primers or working with samples from organisms with very small or no introns may require more complete removal of residual contaminating DNA. The PureLink DNase Set allows for convenient on-column digestion of DNA during the RNA isolation protocol. Treating with DNase while “on-column” is easier and allows higher RNA recovery than treating with DNase after the RNA has been isolated. The PureLink DNase Set can also be used to remove residual DNA from RNA that has been previously purified. Both the “on-column” and post-RNA purification workflows are options available with the PureLink DNase Set.

Simplified, non-toxic RNA purification
The PureLink RNA Mini Kit utilizes non-toxic guanidine-isothiocyanate lysis buffer to protect the RNA during the isolation steps. Special RNase-free reagents combined with certified RNase-free silica membranes in a unique column configuration allow for a safe and easy procedure that can typically be completed in less than 20 minutes without the need of hazardous phenol/chloroform extraction, CsCl centrifugation, or LiCl or alcohol precipitation.

The PureLink RNA Mini Kit is recommended for use with the Homogenizer (Cat. No. 12183026), designed to homogenize cell or tissue lysates via centrifugation, prior to nucleic acid purification. The Homogenizer is especially effective for clarifying particulates from plant tissues.

For plant tissues rich in polyphenolics or starch (e.g., pine needles, potato tubers), we recommend using PureLink Plant RNA Reagent (Cat. No. 12322012) together with the PureLink RNA Mini Kit.

Homogenizer Invitrogen™

The Homogenizer is for use with the Ambion™ PureLink™ RNA Mini Kit (Cat. Nos. 12183020, 12183018A, 12183025) to reduce the viscosity of difficult tissue samples between 5 and 30 mg during RNA isolation.

Ambion™ RNase Inhibitor, cloned, 40 U/µL Invitrogen™

Ambion® RNase Inhibitor, a recombinant human protein produced in E. coli, is a potent inhibitor of neutral pancreatic RNase A type enzymes. The mode of inhibition is noncompetitive; the inhibitor tightly binds RNases in a 1:1 ratio. Supplied in one tube containing 10,000 U (40 U/ µL). The enzyme has been shown to inactivate RNases present in many tissues and cell types. Addition of the ribonuclease inhibitor (RI) has been shown to be useful whenever the integrity of RNA must be maintained such as in the preparation of cDNA by reverse transcription, in vitro RNA transcription, and in in vitro protein synthesis. RI does not inhibit RNase I, T1, T2, H, U1, U2, or CL3. RI requires a minimum of 1 mM DTT to maintain activity, and has an activity range between pH 5.0 to 8.0 with maximal activity between pH 7.0 and 8.0. Since the mode of inhibition is the formation of a 1:1 complex with RNases, care must be taken to avoid denaturation or oxidation of the ribonuclease inhibitor (e.g. by addition of SDS, urea, etc.). RI should be added to transcription, translation, and cDNA synthesis reactions to give a final concentration of 1 U/ µL. RNase Inhibitor is rigorously tested for contaminating RNase, exonuclease, endonuclease, and protease activity.

Unit Definition:
One unit is the amount of protein required to inhibit the activity of 5 ng of RNase A by 50%. Unit assay conditions: 100 mM Tris-acetate (pH 6.5), 1 mM EDTA, 1 mM cyclic 2',3'-CMP, and RNase inhibitor. Addition of RNase A initiates the reaction. RNase Inhibitor activity is measured by the inhibition of hydrolysis of cyclic 2', 3'-CMP by RNase A.

TRIzol™ Plus RNA Purification Kit Invitrogen™

The TRIzol® Plus RNA Purification Kit provides a simple, reliable, and rapid method for isolating high-quality total RNA from a wide variety of samples, including animal and plant cells, tissue, bacteria, and yeast. The kit combines the strong lysis capability of TRIzol® Reagent, followed by a convenient and time-saving silica-cartridge purification protocol from the PureLink® RNA Mini Kit, to provide ultrapure total RNA typically within an hour, even from difficult samples such as fibrous or fatty tissues.

Features of this kit include:

• Combined TRIzol®-based lysis and PureLink™ spin column-based RNA isolation technologies
• Superior lysis capability and convenient protocol

Two products in one
The TRIzol® Plus RNA Purification System couples the lysis capabilities of TRIzol® Reagent with the 1000 µg column-;binding capacity of the PureLink™ RNA Mini Kit, resulting in high RNA integrity and yield. The TRIzol® Plus RNA Purification System includes protocols for a variety of sample types, including fibrous or fatty tissues and plant material. The combination of TRIzol® Reagent followed by silica-cartridge purification of RNA is the recommended RNA purification method in the gene expression industry. The TRIzol® Plus RNA Purification Kit provides the reagents and an optimized protocol to purify total RNA for gene expression studies using an industry recommended method.

Simple lysis to column purification protocol
TRIzol® Reagent sample lysis maintains the integrity of the RNA, while disrupting cells and dissolving cell components. TRIzol® Reagent also provides an immediate and highly effective inhibition of RNase activity during sample homogenization. Following lysis, RNA in the lysed sample is bound to the clear silica-based membrane in the PureLink™ RNA Mini Kit Spin Cartridge, washed to remove contaminants, and then eluted in RNase-Free water (Tris Buffer, pH 7.5 may also be used).

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

MICROBEnrich™ Kit Invitrogen™

For the separation and isolation of bacterial RNA from mixed samples. The Ambion® kit includes sufficient reagents for depleting 500 µg of host cell RNA (20 reactions x 25 µg RNA) from a complex host-bacterial RNA mixture. Gene expression analysis following host-bacterial interactions is revolutionizing our understanding of the host response to bacterial infection. Numerous studies have utilized microarray analysis to follow host cell transcriptome alterations in response to interactions with bacteria. However, because no method has existed to isolate bacterial RNA away from host cell RNA, the corresponding analyses of bacterial transcriptomes has been impossible. The MICROBEnrich™ Kit enables whole genome microarray expression analyses of bacterial RNA after host-bacterial interactions.

Start With Any Total RNA Sample
For microarray analysis and other sensitive applications, a DNase treatment is recommended to remove contaminating genomic DNA from the total RNA prior to the MICROBEnrich™ Kit procedure.

Remove >90% of Mammalian RNA from Complex Total RNA Mixtures
The MICROBEnrichKituses hybridization capture technology (patent pending) to remove human, mouse, and rat RNA (both mRNA and rRNA) from complex host-bacterial RNA populations, leaving behind enriched microbial total RNA. In the first step of the procedure, host-bacterial total RNA is incubated with a mixture of capture oligonucleotides that bind the mammalian 18S and 28S rRNAs and polyadenylated RNAs. Next, the rRNA/oligo nucleotide hybrids and all polyadenylated mRNAs are removed from the mixture with oligonucleotide-derivatized magnetic beads. After magnetic separation, the bacterial RNA remaining in the supernatant and can be precipitated with ethanol. An Agilent 2100 bioanalyzer electropherogram (see Figure) shows nearly complete removal (>95%) of the host 18S and 28S rRNAs from a mixed sample. Bacterial mRNAs purified with the MICROBEnrich™ Kit are better templates than total RNA for downstream applications and therefore deliver dramatic increases in sensitivity when used in applications such as array analysis (see Figure).

The MICROBEnrich™ Kit Will Work With Any Bacterial Species
The MICROBEnrich Kit was optimized using E. coli RNA in a background of human, mouse, and rat RNA. However, the kit should perform well with RNA from any bacterial species. Although many mammalian host species may be compatible with the MICROBEnrichKit, only human, mouse, and rat rRNA sequences were used for the design and optimization of the capture oligonucleotides.

Accessory Products:
A magnetic stand is required for use this kit. They are available in several formats, including the Single Place Magnetic Stand (SKU #AM10026), 6 Tube Magnetic Stand (SKU #AM10055), and 96-well Magnetic Stands (SKU #AM10027 and #AM10050). The RiboPure™-Bacteria Kit (SKU #AM1925) is ideal for the purification of host-bacterial cell total RNA. The TURBO™ DNA-free™ or DNA-free™ DNase Treatment and Removal Reagents (SKU #AM1907 and AM1906, respectively) are recommended for microarray analysis and other applications where the removal of contaminating genomic DNA is needed.After the MICROBEnrich™ procedure, bacterial mRNA can be purified from the total RNA using the MICROBExpress™ Bacterial mRNA Enrichment Kit (SKU #AM1905). This technology enables >95% removal of prokaryotic 16S and 23S rRNAs.

RiboMinus™ Eukaryote Kit for RNA-Seq Invitrogen™

The RiboMinus™ Eukaryote Kit for RNA-Seq provides truly complete transcriptome isolation for next generation sequencing analysis by offering a method for selective depletion of up to 99% of the 5S, 5.8S, 18S and 28S ribosomal RNA from total RNA versus poly-A or cap binding methods, which can introduce bias and select for only part of the transcriptome.

The RiboMinus™ Eukaryote Kit for RNA-Seq Offers:

• A simple, yet selective depletion procedure that uses a sequence-specific probe to capture rRNA
• Rapid and efficient removal of rRNA-probe complex using magnetic bead technology

Three Steps to an rRNA-free Preparation Suitable for Transcriptome Analysis
The removal of large ribosomal RNA using the RiboMinus™ kit is a reliable and robust three-step procedure. Hybridize the RNA sample to the RiboMinus™ Probe in a water bath or heat block, then remove probe-rRNA complexes using the RiboMinus™ magnetic beads included in the kit. Concentrate the depleted RNA preparation by ethanol precipitation, or by using the RiboMinus™ Concentration Module.

Wash Solution II, RNA Purification Invitrogen™

Applied Biosystems® replacement solution for use with TransPrep Chemistry is for the isolation on the ABI PRISM® 6100 Nucleic Acid PrepSation.

Cells-to-CT Bulk RT Reagents Invitrogen™

Cells-to-CT Bulk RT Reagents are separately-available, large-quantity versions of the reagents used for reverse transcription in the TaqMan® Gene Expression 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.

Leukemia (K-562) Total RNA Invitrogen™

The highest-quality RNA available from Ambion®, it is DNase-treated and subjected to unsurpassed quality control standards. One tube containing 100 µg is provided at a concentration of 1 mg/mL. Intactness of mRNA, genomic DNA contamination, presence/absence of enzymatic inhibitors, and ribosomal RNA contamination in poly(A) preparations are all important aspects used to judge the quality of FirstChoice® RNAs. The samples are processed using Ambion® RNA isolation reagents to produce highly pure, intact RNA. A stringent DNase treatment is performed to ensure that the RNA is ready for use in any downstream application, including RT-PCR. Note: While 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. The integrity of the RNA is verified by capillary electrophoresis using the Agilent® 2100 bioanalyzer.

SUPERase•In™ RNase Inhibitor (20 U/μL) Invitrogen™

SUPERase•In RNase Inhibitor is a protein-based inhibitor of non-human origin that noncovalently binds and inhibits the most common and troublesome RNases, including RNase A, B, C, 1, and T1.

SUPERase•In RNase Inhibitor can be used in any application where RNase contamination could be problematic. Because it inhibits a broader range of RNases than traditional RNase inhibitors, SUPERase•In is the most effective RNase inhibitor available, providing a higher level of protection against degradation.

SUPERase•In does not interfere with other enzymes such as RNA polymerases, reverse transcriptase, or Taq DNA polymerase. Additionally, SUPERase•In is active up to 65°C and over a pH range of 5.5 to 8.5.

If you are plannning to use this product with standard antibody purification methods, please contact our technical support to discuss experimental design.

Unit definition
SUPERase•In RNase Inhibitor at 1 U/µL will block the degradation of 0.1 µg/µL labeled RNA by 2.5 pg/µL of RNase A, 2.5 pg/µL of RNase I, and 0.0075 U/µL of RNase T1, for 4 hours at 37°C, in 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 1 mM EDTA. Analysis is by denaturing PAGE. SUPERase•In is currently the only ribonuclease inhibitor for which the unit activity is defined by such a functional assay.

Single Cell Lysis Kit Invitrogen™

Single cell analysis has the potential to transform life science research. However, detecting and quantitating the minute amounts of RNA contained in single cells remains a significant challenge. The Ambion® Single Cell Lysis Kit enables reliable RNA preparation for single cells.

Key product features:

• Optimized sample prep—Enables RNA preparation for single cells
• Efficient design—No sample loss ensures maximal sensitivity
• Versatile application—Compatible with common enzymatic reactions

Optimized RNA Sample Prep for 1-10 Cells
The Ambion® Single Cell Lysis Kit enables RNA preparation from 1-10 cells. The Single Cell Lysis kit generates a concentrated cell lysate through the optimized combination of lysis buffer, stop solution, and DNase. The resulting lysate is compatible with common enzymatic reactions such as reverse transcription. Sample preparation is fast and can be completed in only 7 minutes with 2 pipetting steps (Figure 1).

No Sample Loss Ensures Maximal Sensitivity
The Ambion® Single Cell Lysis Kit bypasses the need for traditional RNA purification. There are no sample transfers, columns or washes which can lead to loss and⁄or dilution of the precious sample. The entire lysate can be used in downstream reactions ensuring maximal sensitivity and accuracy in your experiments.

Compatible with Common Enzymatic Reactions
The reagents in the Ambion® Single Cell Lysis Kit are identical to the lysis reagents in the complete Single Cell-to-CT™ Kit for Real-time RT-PCR analysis from 1-10 cells. They have been validated with multiple reverse transcription kits including SuperScript® VILO™ cDNA Synthesis Kit, High Capacity RNA-to-cDNA kit and TaqMan® MicroRNA Reverse Transcription Kit, for analysis of mRNA as well as miRNA targets. As a stand alone module, the Single Cell Lysis Kit allows you to create new workflows to expand the possibilities of single cell analysis.

PureLink™ Plant RNA Reagent Invitrogen™

Plant RNA Reagent is a single-component solution optimized for the isolation of high yields of high-quality total RNA from a variety of plant tissues, especially those rich in polyphenolics (e.g., conifer needles) or starch (e.g., potato tuber or seeds). Features of the Plant RNA Reagent:

• Isolate high-quality RNA from difficult plant materials
• Use for small-scale and large-scale RNA isolations

Improved quality and purity of plant RNA
Plant RNA Reagent is optimized to have less sample DNA contamination, to be twice as effective as TRIzol® Reagent, and have a simplified RNA extraction protocol for minimized time and labor. The end result is high-yield, high-purity RNA (determined by A260/A280 and gel analysis) from a variety of plant material sources.

Works for milligram to gram quantities of starting material
Plant RNA Reagent provides greater flexibility for sample size and processing formats, and can be used for both small-scale (up to 0.1 g plant tissue) and large-scale (0.1–5 g plant tissue) total RNA isolation. The amount supplied (100 mL) is sufficient for 200 reactions using 100 mg of tissue, or 4 reactions using 5 g of tissue.

Isolated RNA is ideal for a variety of applications and products:

cDNA library construction kits
Real-Time PCR (qPCR)
Northern blotting kits & reagents
Poly(A) selection kit
RNase control reagents & assays

Power SYBR™ Green Cells-to-CT™ Kit Invitrogen™

Ambion® Power SYBR® Green Cells-to-CT™ Kit is a fast, easy, and robust method to go directly from cultured cells to real-time PCR results. A breakthrough cell lysis and RNA stabilization technology eliminates the laborious and time-consuming RNA purification process completely. The kit includes sufficient reagents for 400 reactions. Power SYBR® Green Cells-to-CT™ Kit is Powered by AmpliTaq Gold® DNA Polymerase, LD, which provides the highest levels of specificity with standard real-time PCR.

• Includes optimized reagents to work efficiently, right out of the box
• Simple, effective Cells-to-CT™ technology enables sample preparation at room temperature in only 7 minutes, including DNase treatment
• Validated accuracy, exceptional reproducibility, and maximal sensitivity from 10 to 100,000 cells per sample; results equivalent to those from purified RNA

Extraordinary Value
The kit includes reagents for sample preparation integrated into a complete, optimized gene expression workflow including reverse transcription reagents and a high-performance PCR master mix with SYBR® Green.

Extraordinary Ease
The protocol begins with a simple 7-minute sample preparation procedure. First, cells are washed in PBS, and then lysed for 5 minutes at room temperature; DNase treatment can be performed concurrently. The lysis reaction is stopped, and the sample is then ready for reverse transcription or storage at –20°C for up to 5 months. Because samples are processed directly in culture plates (96- or 384-well), sample handling and the potential for sample loss or transfer error are minimized, resulting in higher reproducibility. Following sample preparation, a portion of the cell lysate is added to an RT reaction followed by real-time PCR using the included master mix.

Extraordinary Performance
To demonstrate the performance of the kit, replicate experiments were run using either traditional RNA purification and real-time PCR analysis, or the Power SYBR® Green Cells-to-CT™ Kit. The kit shows equivalent performance to results obtained using purified RNA for analysis of 10 to 100,000 cells. In addition, the sensitivity, efficiency, and dynamic range were shown to be superior to competitor lysate kits. Samples prepared with the kit have been tested for compatibility on various real-time PCR platforms including those from Applied Biosystems®, Roche, and Bio-Rad.

Accessory Product:
The SYBR® Green Cells-to-CT™ Control Kit (SKU# 4402959) contains a Xeno™ RNA control and primer set and is recommended to monitor the efficiency of cell lysis and amplification inhibition in Cells-to-CT™ reactions.

MICROBExpress™ Bacterial mRNA Enrichment Kit Invitrogen™

The MICROBExpress™ Bacterial mRNA Enrichment Kit is for the purification of bacterial mRNA by removing rRNA from total RNA. Sufficient reagents for 20 mRNA purifications, each from 10 µg of total RNA are provided. Features of the MICROBExpress Kit:

• Dramatically increase the sensitivity of array analyses and other procedures
• Removes >95% of 16S and 23S rRNA from bacterial RNA
• Simple procedure takes less than 2 hours
• Works with many Gram positive and Gram negative bacteria

Rapid purification of bacterial mRNA
Isolating mRNA from bacteria has been virtually impossible, until now. The MICROBExpress Kit uses a novel technology to remove >95% of the 16S and 23S rRNA from total RNA of E. coli and other bacterial species. The kit is suitable for mRNA purification from a broad spectrum of Gram positive and Gram negative bacteria. The 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 applications, and cDNA library construction. Efficient removal of 16S and 23S rRNA from bacterial RNA dramatically increases sensitivity in downstream procedures (see figure).

Start with any total RNA sample
This host-bacterial cell total RNA mixture can be obtained by a variety of RNA isolation methods. The MICROBExpressKit specifically removes rRNAs; small RNAs (i.e., tRNA and 5S rRNA) are not removed. Using total RNA that lacks small RNAs as the starting material for MICROBExpressKit produces the highest possible level of mRNA enrichment (see accessory products). In the first step of the MICROBExpress Kit procedure, total RNA is mixed with an optimized set of capture oligonucleotides that bind to the bacterial 16S and 23S rRNAs. Next, the rRNA hybrids are removed from the solution using derivatized magnetic microbeads. The mRNA remains in the supernatant and is recovered by ethanol precipitation.

Accessory products
A magnetic stand is required for use this kit. They are available in several formats, including the Single Place Magnetic Stand (Cat. No. AM10026), 6 Tube Magnetic Stand (Cat. No. AM10055), and 96-well Magnetic Stands (Cat. Nos. AM10027 and AM10050). The RiboPure™-Bacteria Kit (Cat. No. AM1925) is ideal for the purification of total RNA lacking small RNAs. Small RNAs can be removed from total RNA using the MEGAclear™ Kit (Cat. No. AM1908).
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