Shop All DNA⁄RNA Detection & Analysis Kits

Click-iT™ RNA Alexa Fluor™ 594 Imaging Kit Invitrogen™

The Click-iT® RNA Alexa Fluor® 594 Imaging Kit enables detection of global RNA transcription temporally and spatially in cells and tissues (PNAS (2008) 105:15779-84).The ability to detect newly synthesized RNA or changes in RNA levels resulting from disease, environmental damage or drug treatments is an important aspect of toxicological profiling. Utilizing an alkyne-modified nucleoside, 5-ethynyl uridine (EU), and powerful click chemistry, newly synthesized RNA can be detected without the use of radioactivity or antibodies with a simple, two-step procedure. In step one, the alkyne-containing nucleoside is fed to cells or animals and actively incorporated into nascent RNA. The small size of the tag enables efficient incorporation of the modified nucleoside into RNA, but not into DNA. Detection utilizes the chemoselective ligation or “click" reaction between and azide and an alkyne where the modified RNA is detected with a corresponding azide-containing dye. With its dimunitive “footprint", the Click-iT® detection molecule can easily penetrate complex samples and leaves open the possibility of multiplex analyses with other probes, including antibodies for the detection of RNA-interactive proteins for deeper biological insights.

North2South™ Complete Biotin Random Prime Labeling and Detection Kit Thermo Scientific™

The Thermo Scientific™ North2South™ Biotin Random Prime Labeling Kit provides the components and procedure to successfully prepare biotin-labeled DNA probes for use in Southern blotting and other DNA hybridization methods involving streptavidin detection systems.

This kit includes the North2South Chemiluminescent Detection Kit, which combines a novel ECL substrate for horseradish peroxidase (HRP) and optimized hybridization and blocking buffers to ensure consistent, high-sensitivity Southern blot results.

Features of the North2South Chemiluminescent Detection Kit:

• Sensitivity equal to or greater than 32P
• Much faster than DIG detection systems
• Amenable to nearly any DNA or RNA hybridization method
• Optimized hybridization and blocking buffers and protocol steps yield consistent results from experiment to experiment
• High level of light output and consistent low background allows for very short and multiple exposure times using cooled CCD camera systems or film
• Ready-to-use buffers and short process time makes switching from radioactive detection to chemiluminescent detection an easy transition, even for the novice user
• Specific plaques can be picked out of areas that are very dense with signal due to extremely low background

The North2South Hybridization and Detection Kit combines a novel enhanced luminol substrate for horseradish peroxidase (HRP) with optimized pre-made hybridization and blocking buffers to ensure consistent results and high sensitivity. Any nucleic acid blot can be hybridized with the appropriate biotinylated (biotin-labeled) oligonucleotide probe. After a brief membrane blocking step, the biotin label is detected with streptavidin- HRP and a 5-minute incubation with the chemiluminescent substrate. Results are easily recorded by brief exposure to X-ray film (1-15 minutes) or CCD camera.

Related Products
North2South™ Chemiluminescent Hybridization and Detection Kit
North2South™ Chemiluminescent Substrate for HRP
North2South™ Hybridization Buffer
North2South™ Hybridization Stringency Wash Buffer (2X)

TaqMan™ GMO Maize 35S Detection Kit Applied Biosystems™

This kit contains reagents needed to reliably detect the presence of genetically modified organism (GMO)-specific DNA sequences in seed, grain, processed foods and their ingredients.

• Universal assay detects and quantifies all approved genetically modified organisms (GMOs) in Europe and the vast majority of GMOs approved in other countries
• Fluorogenic 5' nuclease assay using TaqMan® probes provides accurate and reproducible quantitative results
• Label license conveys the PCR service rights for GMO testing when used in conjunction with an Authorized Thermal Cycler, so no additional royalties are payable
• Comprehensive controls provide high confidence in results
• Innovative GMO quantitation software allows users to quickly and easily determine the percentage of GMO content
• Offers the complete solution including sample preparation, instruments, reagents, analysis software, technical support, instrument service, and GMO expertise

The Complete Solution
The TaqMan® GMO Detection and Quantitation Kit is the first of a full product line of GMO assays in development that can detect the presence of GMO-specific DNA sequences in seed, grain, and processed foods and their ingredients. The TaqMan® GMO kit is part of a highly sophisticated but user-friendly system that includes DNA sample preparation, automated PCR amplification and signal detection, data analysis, and quantitation software, thus eliminating the need for personnel trained in molecular techniques.

Reliable TaqMan® Chemistry
The system uses the patented fluorogenic 5' nuclease method for multiplex detection and quantitation, the latest innovation in real-time PCR technology. It targets the Cauliflower Mosaic Virus 35S promoter, a GMO-specific sequence present in all GMO soy and maize events approved for use in food by the European Union, and in the vast majority of GMO events approved by other countries. Both the European Union and Japan view this method as the premier technology for quantitating GMO content in food. Other TaqMan® kits are available for the detection and quantitation of food-borne pathogens, including Salmonella and E. coli 0157:H7.

For Research Use Only. Not for use in diagnostics procedures.

Qubit™ RNA IQ Assay Kit Invitrogen™

The Qubit RNA IQ Assay Kit provides a fast, simple method to check whether an RNA sample has degraded using the Qubit 4 Fluorometer (required). The assay utilizes two unique dyes: one that binds to large, intact and/or highly structured RNA (mRNA, tRNA, rRNA), and another that selectively binds to small and/or, degraded RNA. Together they enable you to quickly assess the quality and integrity of an RNA sample.

To use, simply add your samples to the RNA IQ working solution, then measure on the Qubit 4 Fluorometer. Results are presented as an RNA IQ number (RNA IQ#) that indicates the RNA sample integrity and quality, and also as a calculated percent of large and small RNA in the sample. The RNA IQ# is a value from 1 to 10, similar to other RNA quality scores, where a small number indicates that the sample is comprised of mainly small RNA and a larger number indicates that the sample consists of mainly large RNA or RNA with tertiary structure.

Solution-based compared to electrophoresis methods
The RNA IQ assay results correlate well with electrophoretic methods. The primary difference between the solution-based RNA IQ assay and traditional electrophoresis-based techniques is the amount of time it takes to ascertain whether an RNA sample has degraded. Gels are considered easy to perform, but still take time to run. Microfluidic-based methods not only take time to obtain results, but often have lengthy procedures to prepare the samples. Gel or microfluidic-based electrophoresis is still recommended to obtain fragment size information or size distribution of the RNA sample.

Notes:
1. The Qubit RNA IQ assay will only work on the Qubit 4 Fluorometer. It will not work on the Qubit, Qubit 2.0, or Qubit 3 fluorometers.
2. 500 µL thin-walled PCR tubes (Cat. No. Q32856) are required but not included.

SOLiD™ ChIP-Seq Kit, with ChIP magnet Applied Biosystems™

The SOLiD™ ChIP Seq Kit is a complete and optimized system for genome wide Chromatin Immunoprecipitation (ChIP) analysis on SOLiD™ and includes ChIP preparation reagents and library generation reagents for 20 samples along with a protocol optimized for library preparation with reduced amount of DNA from 1-10ng samples. Chromatin immunoprecipitation (ChIP) experiments examine histone modifications and genomic DNA sequences bound to specific regulatory proteins and when combined with next generation sequencing is a powerful method to examine genome wide protein DNA interactions. This configuration also includes the DynaMAG®-PCR magnet for processing ChIP samples.

MAXIscript™ T7 Transcription Kit Invitrogen™

The Ambion® MAXIscript® T7 In Vitro Transcription Kit synthesizes RNA probes in just 10 minutes for use in ribonuclease protection assays, in situ hybridization, and blot hybridizations. Each kit contains sufficient reagents for 100 reactions.

Features:

• High yields of high specific activity, full-length RNA probes in just 10 minutes
• Ideal probe synthesis kit for nuclease protection assays, northern and Southern blots, and in situ hybridization
• Includes TURBO DNase™
• All kits contain quality-tested DNase/RNase-free reagents

The MAXIscript® In Vitro Transcription Kit synthesizes RNA probes with specific activities reaching 1 × 109 cpm/µg in just 10 minutes. MAXIscript® Kits are very efficient at synthesizing full-length probes, even at limiting nucleotide concentrations, making them the perfect companion to ribonuclease protection assays. MAXIscript® Kits are also an excellent choice for the synthesis of nonisotopically labeled probes. Labeling can be accomplished post-transcriptionally by using Ambion's BrightStar® Psoralen-Biotin Labeling Kit, or during transcription by incorporating modified nucleotides into the reaction. MAXIscript® Kits can also be used for the synthesis of modest amounts of unlabeled RNA (2–6 µg/20 µl reaction) of up to 2 kb.

MEGAscript™ T7 Transcription Kit Invitrogen™

The MEGAscript® T7 Kit is an ultra-high yield in vitro transcription kit. The high yields are achieved by modifying typical transcription reaction conditions so that very high nucleotide concentrations can be effectively used. The T7 Enzyme Mix and the 10X Reaction Buffer are specifically calibrated for each lot and RNA polymerase.

Using the MEGAscript® T7 Kit
The MEGAscript® T7 Kit contains in vitro transcription reaction components and a control template. The kit will yield a total of 3–5 mg of RNA (approximately 100 µg of RNA or more per reaction) from the control template supplied with the kit. This corresponds to 400–650 moles of RNA for each mole of template. Smaller templates typically yield a lower mass and a higher molar yield of product. Novel transcription reaction conditions and a patented, high-yield technology allow the synthesis of 10–50 times the amount of RNA produced by conventional transcription reactions.

Applications
The MEGAscript® T7 Kit is intended for the synthesis of large amounts of unlabeled or low specific activity RNA for a variety of uses, including in vitro translation, antisense/microinjection studies, and isolation of RNA binding proteins. In large-scale transcription reactions, the concentration of all 4 nucleotides is high, well above the Km for the enzyme. The MEGAscript® T7 Kit typically yields over 10 times more RNA than conventional in vitro transcription reactions. The kit is not recommended for synthesis of high specific activity probes.

Accessory products
MessageAmp™ aRNA amplification kits are powered by MEGAscript® technology and have been developed especially for the amplification of cRNA prior to array analysis. To purify in vitro transcription products from free nucleotides, buffer components, and enzymes, we recommend using the MEGAclear™ Kit (Cat. No. AM1908) and the MEGAclear™-96 Kit (Cat. No. AM1909).

TaqMan™ GMO Soy 35S Detection Kit Applied Biosystems™

This kit contains reagents needed to reliably detect the presence of genetically modified organism (GMO)-specific DNA sequences in seed, grain, processed foods and their ingredients.
• Universal assay detects and quantifies all approved genetically modified organisms (GMOs) in Europe and the vast majority of GMOs approved in other countries.
• Fluorogenic 5' nuclease assay using TaqMan® probes provides accurate and reproducible quantitative results.
• Label license conveys the PCR service rights for GMO testing when used in conjunction with an Authorized Thermal Cycler, so no additional royalties are payable.
• Comprehensive controls provide high confidence in results.
• Innovative GMO quantitation software allows users to quickly and easily determine the percentage of GMO content.
• Offers the complete solution including sample preparation, instruments, reagents, analysis software, technical support, instrument service, and GMO expertise.

The Complete Solution
The TaqMan® GMO Detection and Quantitation Kit is the first of a full product line of GMO assays in development that can detect the presence of GMO-specific DNA sequences in seed, grain, and processed foods and their ingredients. The TaqMan® GMO kit is part of a highly sophisticated but user-friendly system that includes DNA sample preparation, automated PCR amplification and signal detection, data analysis, and quantitation software, thus eliminating the need for personnel trained in molecular techniques.

Reliable TaqMan® Chemistry
The system uses the patented fluorogenic 5' nuclease method for multiplex detection and quantitation, the latest innovation in real-time PCR technology. It targets the Cauliflower Mosaic Virus 35S promoter, a GMO-specific sequence present in all GMO soy and maize events approved for use in food by the European Union, and in the vast majority of GMO events approved by other countries. Both the European Union and Japan view this method as the premier technology for quantitating GMO content in food. Other TaqMan® kits are available for the detection and quantitation of food-borne pathogens, including Salmonella and E. coli 0157:H7.

For Research Use Only. Not for use in diagnostics procedures.

TaqMan™ GMO Screening Kit

The TaqMan® GMO Screening Kit analyzes DNA extracted from food and feed samples for the presence of the main transgenic regulatory regions. This real-time PCR kit contains all reagents (except for the template DNA) required for the detection of genetically modified organisms (GMOs ). In addition, the kit contains an Internal Positive Control (IPC) to identify possible PCR inhibition. The benefits of the Taqman® GMO Screening Kit over other commercially available products include:

• Flexibility—GMOs detected in DNA obtained from ANY food or feed source
• Accuracy—an internal positive control (IPC) rules out inhibition during qPCR process
• Speed—results provided in less than 3 hours

The TaqMan® GMO Screening Kit is capable of multiplex amplification of Endogenous Vegetal Control, Internal Positive Control, and these regulatory regions:

• P35S/CaMV
• TNOS/A. tumefaciens
• P34S/FMV

All of which make possible the detection of significant events such as soya MON89788.

The TaqMan® GMO Screening Kit provides all of the necessary reagents for testing 48 samples and is optimized for use in all Applied Biosystems® real-time PCR instruments. And because it is a real-time PCR kit, no electrophoresis is required. In addition, the kit offers extremely high sensitivity, detecting targets with only 3 DNA copies.

The development and manufacture of this kit occurs through a partnership with IMEGEN (Instituto de Medicina Genomica), which has a long history of GMO research and development.

Other related products include:
Taqman® Roundup Ready Soya Quantification Kit
GMO Extraction Kit
TaqMan® GMO Maize Quantification Kit

ViewRNA™ Tissue Assay Blue Module Invitrogen™

This VewRNA Tissue Assay Blue Module is designed for use with the ViewRNA Tissue Assay Core Kit. The Core Kit, when combined with a ViewRNA Tissue probe set (purchased separately), enables the detection of a single mRNA target in tissue sections using the Fast Red substrate. The ViewRNA Tissue Assay Blue Module enables the detection of a second mRNA target using the Fast Blue substrate and must be used in combination with the Core Kit and two probes sets in order to run a 2-plex assay. Each kit contains sufficient reagents to process a minimum of six slides at a time. ViewRNA TYPE 1 probe sets are for use with the Fast Red substrate (Core Kit), while ViewRNA TYPE 6 probe sets are for use with the Fast Blue substrate (Blue Module).

Learn more about and order ViewRNA Tissue probe sets ›

ViewRNA Tissue assays are RNA in situ hybridization (ISH) assays for the reliable detection of one or two mRNA targets within tissue sections. Visualize any gene in any tissue with single-copy sensitivity and no radioactivity. And in case your needs are not met by one of our 6500 catalog probe sets, we have decades of bioinformatics expertise and will design custom probe sets to any mRNA target of interest, free of charge.

With ViewRNA Tissue assays, you can achieve:
• Robust, single mRNA molecule detection in tissue sections
• Accuracy and specificity inherent to branched DNA amplification technologies
• Simultaneous detection of two RNA targets

The ViewRNA Tissue Assay is compatible with formalin-fixed, paraffin-embedded (FFPE) tissue or OCT (optimal cutting temperature) embedded frozen tissue sections. Whereas traditional ISH technologies often amplify both signal and background, ViewRNA assays use branched DNA (bDNA) technology to amplify target-specific signal and achieve superior signal-to-noise ratios.

RapidFinder™ STEC Screening Assay Applied Biosystems™

The Applied Biosystems™ RapidFinder™ STEC Screening Assay is part of a complete, validated screening and confirmation workflow enabling TaqMan™ real-time PCR detection of the Shiga toxin-producing E. coli (STEC) O157:H7 and Big 6 non‐O157:H7 serogroup (O26, O45, O103, O111, O121 and O145) in a variety of beef products. The RapidFinder STEC Screening Assay was co-developed with the USDA-ARS and contains 96 complete qPCR assays targeted to screen for Shiga toxin 1 and 2 (stx1/stx2), eae, and potential O157:H7 presence. The assay is designed to be inclusive of all know allelic variants of stx and eae, including stx2g and stx2f. All reactions include an internal positive control (IPC) and ROX™ passive reference. Samples assessed as negative require no further testing. Samples assessed as positive must be further tested using the RapidFinder™ STEC Confirmation Assay.

The RapidFinder STEC Detection Workflow provides:
• Greater confidence in results—part of a validated workflow for detection of as little as 1-5 CFU of pathogenic STEC in your sample
• Simplified workflow—simultaneous detection of E. coli O157:H7 and the Big 6 non-O157:H7 strains in 375 g beef samples with a closed-tube, single-step lyophilized detection format that fits within your existing testing program
• Reduced chance of false results—through use of high-quality DNA preparation, proprietary assay design, and internal positive control
• Same day detection of pathogenic STECs and clearance of STEC-negative beef samples in as little as 12 hours, supporting early product release
• Minimized costs and laboratory space with the use a lower volume of a nonproprietary single-step enrichment medium, eliminating the need to purchase proprietary media
• Compatibility with RapidFinder™ Express software v.1.2 and later—enables easy, automated presence/absence detection and interpretation of results

Studies have shown that the RapidFinder STEC assays have a high level of sensitivity and specificity for E. coli O157:H7 and the Big 6 non-O157 STEC strains*. They detected all inclusion strains and showed no cross-reactivity to any of the exclusion strains tested. The stx assays detected all known variants of stx1 and stx2, including stx2f and stx2g. The optimized workflow co-developed with USDA-ARS showed equivalent detection to the MLG reference method, and the internal positive control in both assays significantly reduces the risk of false-negatives.

Applications
The RapidFinder STEC Detection Workflow is intended for use by microbiological analysts who need to test for STEC in beef samples. Combined with the Applied Biosystems 7500 Fast RT-PCR System, RapidFinder STEC screening and confirmation assays provide a comprehensive solution for rapid detection, identification, and confirmation of Big 6 STECs and E. coli 0157:H7 to meet importation and exportation requirements for the beef industry. Specific workflows are detailed in the figures below and include both automated and spin column-based DNA isolation and real-time PCR detection of E. coli O157:H7 and Big 6 non-O157 strains in beef samples.

RapidFinder Express Software
When integrated into the RapidFinder STEC Detection Workflow, RapidFinder Express Software automates data interpretation and generates user instructions on how to proceed with a sample. Data displays are available for novice and advanced users, showing simple presence/absence calls or raw data files using 7500 Fast System SDS Software.

RapidFinder STEC Screening and Confirmation Assays meet USDA FSIS guidance
The RapidFinder STEC Detection Workflow, a two-stage real-time PCR method, is designed to accurately detect, identify, and confirm E. coli O157:H7 and the Big 6 serogroups of non-O157 Shiga toxin-producing E. coli defined by the USDA Food Safety and Inspection Service (FSIS) as adulterants in the American beef industry.

Learn more about the RapidFinder STEC Detection Workflow >

*Swimley, Michelle S. Design and Evaluation of Two-Stage Multiplex Real-Time PCR Method for Detecting O157:H7 and non-O157 STEC Strains from Beef Samples. IAFP 2015.

mMESSAGE mMACHINE™ T7 ULTRA Transcription Kit Invitrogen™

The mMESSAGE mMACHINE® T7 Ultra Kit combines a new cap analog —Anti-Reverse Cap Analog (ARCA)—with a patented high-yield transcription technology, to generate RNA transcripts that produce higher protein yields compared to other transcripts upon translation. The kit includes sufficient reagents for 50 reactions. Advantages of the mMESSAGE mMACHINE® T7 Ultra Kit:

• Synthesize more protein from in vitro-transcribed RNA
• Express RNA transcripts more efficiently both in vitro and in vivo
• Stabilize transcripts in vivo with included reagents for poly(A) tailing

ARCA-capped RNA is translationally more active
A base modification in ARCA results in its incorporation in the functional, translatable orientation only; traditional cap analog can be incorporated in both functional and nonfunctional orientations. As a result, incorporating ARCA into transcription reactions yields capped RNAs that are 100% translatable. This is further enhanced by the inclusion of poly(A) tails in mMESSAGE mMACHINE® T7 Ultra transcripts. Experiments comparing ARCA and ARCA/poly(A)–tailed transcripts to cap analog and cap analog/poly(A)–tailed transcripts indicate higher levels of protein synthesis with ARCA capped RNA .

What Is ARCA?
Anti-Reverse Cap Analog (ARCA) is a modified cap analog in which the 3' OH group (closer to m7G) is replaced with OCH3 (see schematic). Because of this substitution, the RNA polymerase can only initiate transcription with the remaining hydroxyl group, thus forcing ARCA incorporation in the forward orientation. As a result, 100% of the transcripts synthesized with ARCA at the 5' end are translatable, leading to a strong stimulatory effect on translation.

Proper capping of in vitro transcribed RNA
Proper capping of RNA promotes correct initiation of protein synthesis, as well as stability and processing of mRNA in vivo. Uncapped RNA is rapidly degraded by cellular RNases after microinjection or transfection into cells. Capped RNA is also typically translated more efficiently in in vitro translation systems, generating RNAs with cap analog incorporated only in the functional orientation. Therefore, substitution of traditional cap analog with ARCA results in the synthesis of capped RNAs that are 100% translatable.

TaqMan™ Roundup Ready Soya Quantification Kit

The TaqMan® Roundup Ready Soya Quantification Kit is a food safety testing kit used to determine the percentage of Roundup Ready soya against the total amount of soya present in a DNA sample. This real-time PCR kit contains all reagents (except for the template DNA) required for the quantification of Roundup Ready soya. The benefits of the TaqMan® Roundup Ready Soya Quantification Kit over other commercially available products include:

• Efficiency—detection of small DNA fragments through use of Taqman® MGB probes
• Sensitivity—quantification limit of 20 DNA copies and detection limit of 3 DNA copies
• Flexibility—adapted to processed DNAs

The TaqMan® Roundup Ready Soya Quantification Kit consists of plasmid-containing targets for the endogenous soy gene, known as lectin, and for the transgenic event (GTS-40-3-2) present in Roundup Ready soy. This soy accounts for approximately 80% of all soy cultivated worldwide and is considered to be the most widespread transgenic event.

The TaqMan® Roundup Ready Soya Quantification Kit is optimized for use in all Applied Biosystems® real-time PCR instruments.

The development and manufacture of this kit occurs through a partnership with IMEGEN (Instituto de Medicina Genomica), which has a long history of GMO research and development.

Other related products include:
GMO Extraction Kit
TaqMan® GMO Screening Kit
TaqMan® GMO Maize Quantification Kit

Pierce™ Magnetic RNA-Protein Pull-Down Kit Thermo Scientific™

The Thermo Scientific Pierce Magnetic RNA-Protein Pull-Down Kit provides researchers with a streamlined, robust method to enrich protein-RNA interactions using end-labeled RNA as the bait.

Features of the Magnetic RNA-Protein Pull-Down Kit:

Direct – capture ribonucleoprotein complexes directly with end-labeled RNA; does not use or require antibodies for pull-down
Easy to use – no spin cups or centrifugation necessary for the enrichment of RBP; procedure streamlined for minimal hands-on time (less than 3 hours) after RNA labeling reaction
Flexible – use in vitro transcribed RNA or synthetic RNA for labeling of various lengths and complexity; proteins successfully enriched using endogenous, over-expressed, and in vitro translated lysates
Specific – low bead background; unrelated RNA or mutated RNA does not significantly enrich specified RBPs
Economical – less expensive than purchasing commercially synthesized end-labeled RNA, magnetic beads, and reagents separately
Complete – contains both labeling and enrichment modules with buffers necessary for assay; positive control RNA, negative control RNA, and RBP antibody included

The Magnetic RNA-Protein Pull-Down Kit provides reagents to efficiently enrich RNA Binding Proteins (RBPs) using RNA end-labeled with desthiobiotin and streptavidin magnetic beads. The complete kit contains sufficient reagents for 20 RNA labeling reactions and 20 protein-RNA pull-down assays. This direct enrichment of the protein-RNA interaction provides an alternative to antibody capture of protein-RNA complexes or moieties incorporated into the nucleic acid. An added advantage to the kit is that it includes validated controls for both the labeling and pull-down assay. The kit is amenable to several downstream applications, including Western blotting and Mass Spectrometry (MS).

Includes:
Complete kit contains the Pierce RNA 3'-End Desthiobiotinylation Kit, positive and negative RNA controls, nucleic-acid compatible streptavidin magnetic beads, and buffers for RBP enrichment and elution

Requires:
User-supplied RNA and lysate (experimental sample)

Applications:
• Mutational analysis
• Structure function analysis
• Identification of RNA:Protein interactions
• MS analysis of unknown RNA:protein binding pairs or complexes

Utilizing labeled RNA as bait for RNA-binding protein enrichment is advantageous over antibody enrichment in that it captures the interaction directly. Included in this kit is the Pierce RNA 3'-End Desthiobiotinylation Kit, which uses T4 RNA ligase to attach a single cytidine bisphosphate nucleotide to the 3'-ends of single-stranded RNA. Desthiobiotin may be used for detection or as an elutable affinity handle. The length of the spacer between the nucleotide and desthiobiotin has been optimized for efficient attachment to the beads without compromising the accessibility of the RNA to protein.

The procedure for enrichment of RNA binding proteins has been optimized for ease-of-use. The labeled RNA is first captured to the beads to orient the RNA for protein binding. RNA-bound beads are then equilibrated in Protein RNA Binding buffer before protein lysate is added. After washing, sample can be eluted using either nondenaturing Biotin Elution Buffer or SDS-PAGE loading buffer. Eluted samples are ready for a variety of downstream applications, including Western blotting or Mass spectrometry.

The control system for the pull-down assay utilizes 3'-untranslated region androgen receptor (AR) RNA, poly(A)25 RNA, and mammalian cell lysate. The proximal 3'-untranslated region (UTR) of androgen receptor RNA contains UC-rich regions for HuR and Poly(C) Binding Proteins (CP1 and 2). These RNA binding proteins regulate mRNA stability (HuR) and mRNA turnover and translation (CP1 and 2). The negative control RNA, poly(A)25 RNA, does not contain HuR or poly(C) BP binding sites.

Related Products
Pierce™ Magnetic RNA-Protein Pull-Down Kit

Biotin Chromogenic Detection Kit Thermo Scientific™

Thermo Scientific Biotin Chromogenic Detection Kit is a convenient tool for the chromogenic detection of biotinylated nucleic acid probes. The kit is optimized to reproducibly provide high sensitivity with a low background in applications, such as Southern, Northern, dot and slot blotting, and screening of viral plaques and bacterial colonies. Biotinylation of DNA/RNA probes is widely used as a safe and convenient alternative to radioactive labeling. Biotin can be incorporated into nucleic acids using various enzymatic or non-enzymatic methods, including the Biotin DecaLabel DNA Labeling Kit.

Features

Biotinylated probes are detected with streptavidin coupled to alkaline phosphatase (AP). Streptavidin-AP conjugates bind specifically and irreversibly to the biotin-labeled probes. The probes are then visualized using a chromogenic substrate for alkaline phosphatase BCIP/NBT, which produces a blue-purple precipitate. Visualization does not require X-ray film or other specific equipment (see Figure 1).

Highlights

High sensitivity of detection—0.03 pg of homologous DNA in dot blot hybridization with biotin-labeled probe
Low background due to optimized washing and blocking procedures
Non-radioactive detection visualization does not require X-ray film or other specific equipment (see Figure 2)
Fast and convenient—ready-to-use components

Applications

• Southern, Northern, dot/slot blots, plaque, or colony screening.

Note:This kit is optimized for hybridization procedures using nylon membranes. The detection sensitivity may vary depending on the membranes supplied by different vendors.

Related Products
Biotin Chromogenic Detection Kit
Results per page
    spinner