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MagMAX™-96 AI/ND Viral RNA Isolation Kit (Applied Biosystems™)

Applied Biosystems® This kit is for the isolation of viral RNA and DNA from diverse sample types (i.e., environmental samples and biofluids). Nucleic acids purified using this kit are ideal for PCR and RT-PCR applications. This is a first generation MagMAX™ Viral RNA Isolation Kit validated by the NVSL for use with AI/ND viruses. Perfect for working with low viral concentrations, the kit contains sufficient reagents for 4 x 96 reactions.

• Validated by the NVSL for use with AI/ND viruses
• Recover as few as 10 copies of viral RNA from 100–400 µL sample
• Typical viral RNA recovery of >50%
• Ideal when working with low viral concentrations
• Linear recovery down to 50 transcripts
• 96 samples can be processed in 30 minutes
• Easily integrated into robotic platforms

Benefits of MagMAX™ Magnetic Bead-Based Purification:
Magnetic beads offer many benefits compared to other technologies for isolating RNA. Beads bind RNA more efficiently than glass fiber filters, resulting in higher and more consistent RNA yields. Additionally, because filters are not used, there is no risk of filter clogging due to cellular particulates in samples. Since only a small volume of magnetic beads is needed for high-efficiency binding, the bound RNA can be eluted in just 20–50 µL of nuclease-free water, concentrating RNA from large, dilute samples.

Optimized for Isolation of Virus from Biofluids:
The Applied Biosystems® MagMAX™ Viral RNA Isolation Kit has been optimized for use with biological fluids and cell-free samples such as serum, plasma, swabs, and cell culture media. These samples often present unique challenges when purifying RNA, including low viral titers, large volumes to be processed, and sample complexity. Without the need for organic solvents or RNA precipitation, the MagMAX™ Viral RNA isolation kit eliminates the problems often encountered with other RNA isolation methods such filter clogging, large elution volumes, and inconsistent yields.

Get the Same Amount of RNA from 10-fold Less Sample
The sensitivity of viral RNA detection was compared using RNA isolated with Applied Biosystems® MagMAX™ protocol or the current National Veterinary Services Laboratories (NVSL) validated viral testing protocol. Even at one-tenth the sample input, viral RNA isolated using the MagMAX™ kit could be detected at equal or greater levels than RNA isolated using the NVSL protocol and recommended competitor's kit (see data). Downloadable protocols for the use of this kit with specific liquid handling systems are available at our Automation Resource.

Accessory Products:
Carrier RNA (SKU#4382878), Lysis/Binding Solution Concentrate (SKU#AM8500), Wash Solution 1 Concentrate (SKU#AM8504), and Wash Solution 2 Concentrate (SKU#AM8640) are components of the MagMAX™ Viral RNA Isolation Kits that may also be purchased separately. A 96-well magnetic ring stand is required to use this kit. The Magnetic Stand-96 (SKU#AM10027) and the 96-well Magnetic Ring Stand (SKU#AM10050) are designed for paramagnetic bead precipitation from standard 96 well U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors. For additional processing plates, Matrix 96-Well U Bottom Polypropylene Plates (Thermo Scientific, Item# 4917) are recommended. The MagMAX™ and MagMAX™-96 Viral RNA Isolation Kits (SKU#AM1939 and SKU#AM1836) are the newest generation MagMAX™ kits for this application, but are not yet validated by the NVSL.

MagMAX™-96 Viral RNA Isolation Kit (Applied Biosystems™)

The MagMAX-96 Viral RNA Isolation Kit is for the isolation of viral RNA and DNA from diverse sample types (e.g., environmental samples and biofluids). Nucleic acids purified using this kit are ideal for PCR and RT-PCR applications. For a NVSL-validated version of this kit, please see the MagMAX-96 AI⁄ND Viral RNA Isolation Kit.

Benefits of MagMAX magnetic bead-based purification:
Magnetic beads offer many benefits compared to other technologies for isolating RNA. Beads bind RNA more efficiently than glass fiber filters, resulting in higher and more consistent RNA yields. Additionally, because filters are not used, there is no risk of filter clogging due to cellular particulates in samples. Since only a small volume of magnetic beads is needed for high-efficiency binding, the bound RNA can be eluted in just 20–50 µL of nuclease-free water, concentrating RNA from large, dilute samples.

Optimized for isolation of virus from biofluids
The MagMAX Viral RNA Isolation Kit has been optimized for use with biological fluids and cell-free samples such as serum, plasma, swabs, and cell culture media. These samples often present unique challenges when purifying RNA, including low viral titers, large volumes to be processed, and sample complexity. Without the need for organic solvents or RNA precipitation, the MagMAX Viral RNA Isolation Kit eliminates the problems often encountered with other RNA isolation methods such filter clogging, large elution volumes, and inconsistent yields. With this kit, DNA and RNA input levels ranging from 20 copies to 25 x 106 copies have been quantitatively recovered from plasma, serum, and milk. And even with less sample input, pathogen detection was as sensitive (equivalent CTs) or more sensitive (lower CTs) using nucleic acid isolated with the MagMAX kit when compared to nucleic acid isolated using another company's product.

Accessory Products:
This kit is optimized for automated nucleic acid purification using KingFisher magnetic particle processors. This kit can also be used for manual nucleic acid purification using a 96-well magnetic ring stand. The Magnetic Stand-96 (Cat. No. AM10027) and the 96-well Magnetic Ring Stand (Cat. No. AM10050) are designed for paramagnetic bead precipitation from standard 96-well, U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors.

MagMAX™-96 Blood RNA Isolation Kit (Invitrogen™)

This Ambion® kit is for the rapid, high-throughput purification of pathogen RNA from mammalian whole blood, milk, and other biofluids in 96-well plates. RNA purified using the kit is ideal for RT-PCR applications. The kit uses magnetic bead–based purification and includes sufficient reagents for 96 reactions.

• Easily isolate viral or total RNA from whole blood
• Save precious time—complete the protocol in less than 1 hour
• Walk away—integrate with established robotic platforms
• Validated with human, bovine, porcine, and ovine whole blood

Benefits of MagMAX™ Magnetic Bead-Based Purification:
Magnetic beads offer many benefits compared to other technologies for isolating RNA. Beads bind RNA more efficiently than glass fiber filters, resulting in higher and more consistent RNA yields. Additionally, because filters are not used, there is no risk of filter clogging due to cellular particulates in samples. Since only a small volume of magnetic beads is needed for high- efficiency binding, the bound RNA can be eluted in just 30–50 µL of nuclease-free water, concentrating RNA from large, dilute samples.

Isolate RNA from as Little as 50 µl of Blood:
For the highest-quality RNA, use whole blood samples that have not been frozen. Approximately 300–500 ng of intact RNA can be consistently purified from 50 µl of human whole blood. Bovine, porcine, and ovine whole blood samples can also be used. Yields vary from species to species (see data). The recovered RNA is devoid of PCR inhibitors such as proteins and heme, commonly found in RNA isolated from blood.

Isolate Viral and Total RNA from Whole Blood Samples:
The MagMAX™-96 Blood RNA Isolation Kit eliminates sample preprocessing, which can cause sample mix-up and cross-contamination. The procedure is fast (<1 hour), simple, and well suited for automation. The kit can also be used to process fewer than 96 samples at one time and is compatible with either manual processing using multi-channel pipettors or processing with robotic liquid handlers. Downloadable protocols for the use of this kit with specific liquid handling systems are available at our Automation Resource. The MagMAX™-96 Blood RNA Isolation Kit can also be used for total nucleic acid isolation by omitting the DNase treatment.

Accessory Products:
Lysis/Binding Solution Concentrate (SKU#AM8500), Wash Solution 1 Concentrate (SKU#AM8504), and Wash Solution 2 Concentrate (SKU#AM8640) are components of the kit that may also be purchased separately. A 96-well magnetic ring stand is required to use this kit. The Magnetic Stand-96 (SKU#AM10027) and the 96-well Magnetic Ring Stand (SKU#AM10050) are designed for paramagnetic bead precipitation from standard 96-well U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors. For additional processing plates, Matrix 96-Well U Bottom Polypropylene Plates (Thermo Scientific, Item# 4917) are recommended.

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.

PureLink™ Quick Gel Extraction and PCR Purification Combo Kit (Invitrogen™)

PureLink™ Quick Gel Extraction & PCR Purification Combo Kit offers the ability to perform both a gel extraction or a PCR purification a single kit.
The PureLink™ Quick Gel Extraction and PCR Purification Combo Kit is designed to purify DNA fragments from agarose gels. The simple procedure uses a silica-based spin cartridge to purify DNA fragments from 40 bp to 10 kb in < 30 minutes. It also provide all components to perform a rapid and efficient removal of short primers, dNTPs, enzymes, short-failed PCR products, and salts from PCR fragments >100 bp in less than 10 minutes. Isolated DNA using the PureLink™ kit is free of proteins, dye, and agarose, and is ready-to-use for a variety of applications, including automated fluorescent DNA sequencing, manual DNA sequencing, PCR, in vitro transcription, restriction mapping, cloning, and labeling. The kit contains all the buffers from the PureLink™ Quick Gel extraction kit and the PCR binding buffer from PureLink™ PCR Purification kit with Invitrogen branded lidded column.

MagMAX™-96 for Microarrays Total RNA Isolation Kit (Invitrogen™)

The MagMAX™-96 for Microarrays Total RNA Isolation Kit is for the rapid, high-throughput purification of RNA from mammalian cells and tissue in 96-well plates. The kit combines an organic extraction with magnetic bead–based purification yielding extremely high-quality RNA suitable for the most stringent applications, such as microarray analysis. The kit includes sufficient reagents for 96 reactions. Advantages of the MagMAX™-96 for Microarrays Total RNA Isolation Kit:

• Improve gene expression studies with consistent results from experiment to experiment
• Combines TRI Reagent® with a superior magnetic bead–based technology
• Streamlined RNA purification, without sacrificing RNA quality or quantity
• Requires less hands-on time than competitor kits
• Can be integrated with established robotic platforms
• Best MagMAX™ kit for difficult samples or large volumes

Benefits of MagMAX™ magnetic bead–based purification
Magnetic beads offer many benefits compared to other technologies for isolating RNA. Beads bind RNA more efficiently than glass fiber filters, resulting in higher and more consistent RNA yields. Additionally, because filters are not used, there is no risk of filter clogging due to cellular particulates in samples. Since only a small volume of magnetic beads is needed for high- efficiency binding, the bound RNA can be eluted in just 20–50 µL of nuclease-free water, concentrating RNA from large, dilute samples.

Highly stringent for the best-quality RNA
The MagMAX™-96 for Microarrays Total RNA Isolation Kit (patent pending) procedure combines the robust and reliable lysis/denaturant TRI Reagent® with MagMAX™ magnetic bead–based RNA purification technology, resulting in streamlined RNA purification, even in a high-throughput format, without sacrificing RNA quantity or quality. TRI Reagent® readily lyses even difficult cells and tissues while denaturing nucleases to maintain the integrity of RNA. The kit provides high yields of pure, intact RNA that can be used directly for quantitative reverse transcriptase PCR (qRT-PCR) and microarray analysis. See the accompanying data showing RNA yield and quality obtained when using the MagMAX™-96 for Microarrays Total RNA Isolation Kit. Also shown is data illustrating a high correlation between the method that Affymetrix recommends for producing RNA for use on their GeneChips® and the method that the MagMAX™-96 for Microarrays kit employs.

Fast, flexible procedure
Two alternative protocols are supplied for the kit. The "Spin" method has fewer steps, requires no DNase treatment, and allows for larger sample amounts to be processed. The "No Spin" method requires less sample handling and can easily be integrated with a robotic liquid handler. The procedure is fast (<1 hr), simple, and well suited for automation. The kit can also be used to process fewer than 96 samples at one time and is compatible with either manual processing using multi-channel pipettors or processing with robotic liquid handlers. Downloadable protocols for the use of this kit with specific liquid handling systems are available at our Automation Resource.

Accessory products
A 96-well magnetic ring stand is required to use this kit. The Magnetic Stand-96 (Cat. No. AM10027) and the 96-well Magnetic Ring Stand (Cat. No. AM10050) are designed for paramagnetic bead precipitation from standard 96-well, U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors. For additional processing plates, Matrix 96-Well U Bottom Polypropylene Plates (Thermo Scientific, Item# 4917) are recommended.

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.

MagMAX™ Total Nucleic Acid Isolation Kit (Applied Biosystems™)

The MagMAX™ Total Nucleic Acid Isolation Kit is designed for the rapid, high-throuphput purification of RNA and genomic DNA from a variety of samples including viral, blood, and bacterial samples. It includes zirconia beads for the mechanical disruption of difficult pathogens. RNA purified using the kit is ideal for PCR and RT-PCR applications. The kit uses magnetic bead–based purification and includes sufficient reagents for 100 reactions.

• Complete, ready-to-use reagents for RNA and DNA purification from tough-to-lyse bacteria
• Highly effective sample homogenization using bead beating and chemical lysis
• Magnetic bead–based purification technology allows effective removal of PCR inhibitors
• Nucleic acid purification is easily scalable and suitable for automation

Proven with a Wide Range of Samples:
• Biological Samples
• Feces and whole blood
• Biofluids such as milk, urine, nasal fluids, swab samples, semen, sputum, and spent culture medium
• Bacterial cultures and mammalian cell cultures
• Environmental Samples
• River and pond water
• Agricultural samples such as manure pits and cow alleyways

Benefits of the MagMAX™ Total Nucleic Acid Isolation Kit:
The MagMAX™ Total Nucleic Acid Isolation Kit procedure combines an effective method sample disruption with magnetic bead–based nucleic acid isolation and purification technology. The MagMAX™ microspherical paramagnetic beads have a large available binding surface and can be fully dispersed in solution, allowing thorough nucleic acid binding, washing, and elution. This results in consistent yields of high-quality nucleic acid with >50% recovery and little sample-to-sample variation (recovery may vary depending on sample type). The entire procedure can be completed in approximately 1 hour.

Overview of the Procedure:
Samples are disrupted using zirconia beads in a guanidinium thiocyanate–based solution; this rapidly releases nucleic acid while simultaneously inactivating nucleases. Samples are then diluted with isopropanol, and paramagnetic beads are added to the sample. The beads bind nucleic acids and are then immobilized on magnets and washed to remove proteins and other contaminants. A second wash solution is used to remove residual binding solution. Finally, nucleic acid is eluted in a small volume of low-salt buffer. The MagMAX™ Total Nucleic Acid Isolation Kit procedure is designed for processing samples in 96-well plates, thus 96 samples can be processed at once; however, the kit can also be used with fewer than 96 samples.

Accessory Products:
Lysis/Binding Solution Concentrate (SKU#AM8500), Wash Solution 1 Concentrate (SKU#AM8504), and Wash Solution 2 Concentrate (SKU#AM8640) are components of the MagMAX™ Total Nucleic Acid Isolation Kit that may also be purchased separately.

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

This Wash Buffer B is included in the following Dynabeads™ mRNA purification kits: Cat. Nos. 61006, 61011, 61012, and 61021. It is made available separately for applications that require more Wash Buffer B than is provided in the kit.

Dynabeads™ mRNA Purification Kit (for mRNA purification from total RNA preps) (Invitrogen™)

The Dynabeads® mRNA Purification Kit rapidly isolates the mRNA transcriptome in typically 15 minutes, delivering pure, intact mRNA. The kit is designed to specifically target, capture, and purify mRNA molecules from total RNA preparations. Advantages of using the Dynabeads® mRNA Purification Kit:

• Obtain pure mRNA in typically 15 minutes
• Recover and enrich the transcriptome efficiently
• Prepare mRNA that is suitable for nearly every downstream application

Pure, rapid mRNA isolation
The Dynabeads® mRNA Purification Kit contains magnetic beads for the isolation of the mRNA transcriptome from any total eukaryotic RNA preparation (see figure). Dynabeads® magnetic beads are uniform in size and highly mobile in solution (see figure). This enables the bead capture surface to quickly and continuously interact with the entire total RNA sample during the mRNA capture phase. The ribosomal RNA and small RNA molecules (transfer RNA, microRNA, small nucleolar RNA, and small cytoplasmic RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured resulting in cleaner, more sensitive results (see figure). Within about 15 minutes, pure mRNA is isolated and ready for use in downstream applications.

A straightforward enrichment procedure
The Dynabeads® mRNA Purification Kit uses a robust affinity purification principle for the enrichment of polyadenylated mRNA. Superparamagnetic Dynabeads®, coupled to oligo-(dT)25, are first equilibrated with Binding Buffer, and then mixed with purified total RNA. 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. RNA is suitable for all downstream molecular applications, including:

• Gene cloning
• cDNA synthesis, cDNA library construction
• RT-PCR, quantitative RT-PCR
• RPA (Ribonuclease Protection Assay)
• Subtractive hybridization
• Dot/slot hybridization
• Primer extension

MagMAX™ DNA Multi-Sample Ultra Kit (with extra proteinase K and DNA binding beads) (Applied Biosystems™)

Note: This product is identical to the MagMAX™ DNA Multi-Sample Ultra Kit (A25597) except that it comes with additional proteinase K and DNA Binding Beads, which some customers find useful for their specific applications.

The MagMAX™ DNA Multi-Sample Ultra Kit is optimized to isolate genomic DNA (gDNA) from a variety of samples, such as whole blood, buccal cells, saliva, urine, blood cards, mouth rinse, and tissue. gDNA purified with the kit is ideal for qPCR applications on sensitive, scalable platforms such as the QuantStudio™ 12K Flex System with OpenArray® Block (with AccuFill™ System). The magnetic bead–based purification format allows you to easily scale from processing 12 to 500 samples a day, making it the ideal choice for pharmacogenomics (PGx) studies.

Features of the MagMAX™ DNA Multi-Sample Ultra Kit include:
• Streamlined protocols for numerous non-invasive biological samples
• Optimized to provide DNA yields suitable for OpenArray® and Ion AmpliSeq™ applications
• Packaged for high-throughput needs

One Kit for a Diverse Set of Sample Types
Why purchase multiple purification kits to isolate gDNA from different sample types? The MagMAX™ DNA Multi-Sample Ultra Kit eliminates the headache of ordering these extra kits because it has been optimized for use with a variety of biological samples. The kit is designed for high-throughput isolation of gDNA from 1 buccal swab, 50 to 350 µL of whole blood, up to 400 µL urine (without sample concentration), up to 150 µL saliva, 1-5 mL mouth rinse, blood spot cards, and mammalian tissue samples (up to 10 mg). The protocols and magnetic bead format make the kit fully amenable to automation.

*Please note that the Thermo Scientific™ KingFisher™ Flex Magnetic Particle Processor with a 24 Deep-well Head is recommended for sample volumes greater than 50 µL.

Benefits of MagMAX™ Magnetic Bead–based Purification
Magnetic beads offer many benefits compared to other technologies for isolating gDNA. Beads bind gDNA more efficiently than glass fiber filters, resulting in higher and more consistent gDNA yields. Additionally, because filters are not used, there is no risk of filter clogging due to cellular particulates in samples.

Accessory Products
Proteinase K (A25561), DNA Binding Beads (A25562), Wash Solution 1 Concentrate (AM8504), and Wash Solution 2 Concentrate (AM8640) are components of the kit that may also be purchased separately.

MagMAX™ Viral/Pathogen Wash Solution (Applied Biosystems™)

This wash solution is included in the MagMAX Viral/Pathogen Nucleic Acid Isolation kits and MagMAX Microbiome Ultra Nucleic Acid Isolation Kit (Cat. Nos. A42352, A42356, A42357, and A42358). It is made available separately for applications that require more wash solution than is provided in the kit.

RiboPure™ RNA Purification Kit, blood (Invitrogen™)

The RiboPure™-Blood Kit is for the isolation of high-quality RNA directly from whole blood. The kit combines two methods of RNA purification to efficiently remove blood components (protein, heme, genomic DNA, and RNases) that can compromise downstream procedures. The kit includes sufficient reagents for 40 RNA purifications from whole blood samples ranging from 0.3–0.5 mL each. Advantages of the RiboPure™-Blood Kit:

• Purify total RNA directly from whole blood—without prior recovery of WBCs
• Simple, fast procedure—from blood to RNA in less than 30 minutes
• Includes RNAlater® Solution—store blood samples 3 days or more at room temperature
• Includes DNA-free™ Reagents—to ready RNA for RT-PCR and array analysis

Applications
The RiboPure™-Blood Kit produces high yields of pure RNA directly from whole blood, eliminating the need to isolate WBCs prior to RNA purification. RNA recovered using the RiboPure™-Blood Kit can be used in the most challenging applications, including real-time RT-PCR and array analysis. Average yields of RNA are between 2–4 µg/0.5 mL of whole blood (variation is due to differences in WBC numbers between samples).

Stabilize RNA in whole blood
When collecting blood samples for RNA analysis, storage of the samples prior to RNA isolation is often required. The RiboPure™-Blood Kit contains RNAlater® Solution for the stabilization of RNA in whole blood. It also "freezes" the gene expression profile of the cells. Treated samples can be safely stored at ambient temperature for extended periods of time (up to three days or more). Blood samples stored in RNAlater® Solution yield RNA of comparable quality to blood samples processed directly (see figure).

Simple procedure yields highly pure RNA
The RiboPure™-Blood procedure consists of three steps: 1) addition of Lysis Solution directly to RNAlater®-treated or fresh whole blood, 2) initial RNA purification by phenol/chloroform extraction, and 3) final RNA purification on a glass fiber filter. An additional DNase treatment can then be performed with the DNA-free™ Reagents included in the kit. The entire procedure, including DNase treatment, can be completed in less than 1 hour.

Accessory products
The RNA can subsequently be enriched for white blood cell (WBC) mRNA using our GLOBINclear™ Kits to selectively remove globin transcripts. The Mouse RiboPure™-Blood RNA Isolation Kit (Cat. No. AM0951) is also available for the recovery of total RNA—and optionally microRNA—directly from mouse or rat blood. Mouse RiboPure™-Blood includes sample tubes preloaded with RNAlater® Solution to prevent RNA degradation and preserve RNA profiles.

JetQuick™ Blood and Cell Culture DNA Maxiprep Kit (Invitrogen™)

The JetQuick™ Blood and Cell Culture DNA Maxiprep Kit is designed for rapid and efficient purification of genomic DNA (gDNA) from large volumes of blood, body fluids (amniotic fluid, saliva, sperm, lymph), and mammalian cells. It utilizes silica membrane-based technology that allows for isolation of 20-50 kb DNA that is suitable for a wide range of downstream applications, including PCR, restriction enzyme digestion, and Southern blotting.

Key benefits include:
• Large volume processing, up to 10 mL of blood or saliva
• Long double-stranded DNA (20-50 kb) recovery
• Compatible with centrifugation or a vacuum manifold

The maxiprep format enables the processing of up to 10 mL of whole blood or other body fluid. DNA from up to 5 x 108 mammalian cells can be isolated. The purified DNA is ready for downstream analysis.

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.