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microRNA & Noncoding RNA Analysis Using Real-Time PCR
MicroRNAs (miRNAs) are small, noncoding RNA molecules that direct posttranscriptional suppression of gene expression. They have been shown to act as key regulators in many basic biological processes such as development, cell proliferation, differentiation, and the cell cycle. Emerging evidence also implicates miRNAs in the pathogenesis of human diseases such as cancers, metabolic diseases, neurological disorders and infectious diseases.
Common miRNA research applications:
- miRNA profiling—identify differentially expressed mature miRNAs using qPCR in application areas like oncology, metabolic diseases, and autoimmune diseases
- miRNA quantitation—specific quantification of mature miRNAs using qPCR
- miRNA validation—confirm miRNA profiling results using qPCR
- miRNA functional analysis—miRNA mimics and miRNA inhibitors for functional studies
- Noncoding RNA (ncRNA) analysis—reliable detection and quantitation of non-coding human, mouse, or rat transcripts longer than 200 nucleotides.
- Small RNA analysis—detect and measure any small RNA
- pri-miRNA analysis—detect and measure primary miRNAs from human, mouse, or rat species
Applied Biosystems TaqMan MicroRNA Assays are innovative tools for miRNA research-from isolation through discovery, profiling, quantitation, validation and functional analysis. We have developed two methods for sensitive, precise quantification and profiling of miRNA using real-time PCR TaqMan probe-based analysis.
- TaqMan Advanced miRNA Assays use ligation-based universal reverse transcription for an easy streamlined workflow.
- TaqMan MicroRNA Assays — this “classic” version employs target-specific stem-loop reverse transcription primers for 3’ extended templates for highly sensitive detection.
Real-time PCR microRNA workflow:
Isolate RNA
Isolate RNA using methods that preserve RNA integrity and expression profiles
- RNA isolation kits
Innovative Ambion and Invitrogen RNA extraction products - mirVana miRNA Isolation kits
These kits use a rapid procedure to isolate total RNA, including small RNAs such as miRNA, small interfering RNA (siRNA), and small nuclear RNA (snRNA), from tissues and cells. We can address a small number of targets or handle samples that require high throughput. - TaqMan miRNA ABC Purification Kit
For use with the classic TaqMan MicroRNA assays, obtain high quality miRNA for sensitive and specific miRNA expression analysis from difficult sample types including blood, serum, plasma, FFPE, solid tissues, cultured cells, saliva, or urine
Design and optimize primers
Not required for Applied Biosystems TaqMan Assays. These assays have passed our industry leading bioinformatics design criteria.
Reverse Transcription
Convert your RNA into cDNA, over a wide range of RNA concentrations.
TaqMan Advanced miRNA cDNA Synthesis Kit
Provides miR-Amp Universal Amplification with universal primers and master mix to increase the amount of cDNA for each target with accurate detection as few as 60 copies from as little as 1pg of total RNA or 2 μL of serum/plasma. Pre-Amplification Step for TaqMan Advanced miRNA Assays.
Note that a pre-amplification step is performed after the RT. The miR-Amp master mix and miR-Amp primer mix are included in the TaqMan Advanced miRNA cDNA Synthesis Kit.
TaqMan MicroRNA Reverse Transcription Kit
Classic TaqMan microRNA kit for use with the TaqMan MicroRNA Assays to convert miRNA to cDNA (miRNA specific RT). Best for 1-10 targets. Not compatible with TaqMan Advanced miRNA products.
Megaplex RT Primers (For TAC and OpenArray)
Designed to streamline experimental workflows, Megaplex RT Primers provide a single reaction solution when preparing cDNA for real-time PCR analysis on a TaqMan MicroRNA Array.
Megaplex PreAmp Primers
Pre-amplification and reverse transcription primer for use with the classic TaqMan MicroRNA assay (not for use with TaqMan Advanced MicroRNA products) when sensitivity is of the utmost importance, Megaplex PreAmp Primers significantly enhance the detection of miRNAs expressed at low levels, enabling the generation of a comprehensive expression profile using very little sample—as little as 1 ng of input total RNA.
- MegaPlex PreAmp Primers
- Megaplex Primer Pools, Human Pools Set v3.0
- Megaplex Primer Pools, Rodent Pools Set v3.0
Note: When used with - OpenArray, TAC with less than 350ng of RNA and optional for TaqMan Array Cards with greater than 350ng of RNA. For single tube assays you can run pre-amplification with the appropriate protocol.
TaqMan PreAmp Master Mix
Stretch as little as 1 ng of cDNA into 200 real-time PCR reactions for gene expression analysis using TaqMan Gene Expression assays
Amplify cDNA
- TaqMan Advanced miRNA Assays
Enable highly sensitive and specific quantification of mature miRNAs using qPCR. This streamlined workflow includes universal reverse transcription (RT)—one RT step for all miRNA targets. - TaqMan MicroRNA Assays
A simple, two-step protocol includes reverse transcription with a miRNA-specific primer, followed by real-time PCR with TaqMan assays - TaqMan Array miRNA Cards and Plates
Enable highly sensitive and specific quantification of miRNAs in a high throughput format. Both classic and Advanced chemistry cards and plates are available. - Custom TaqMan Advanced miRNA Array Cards
- Custom TaqMan miRNA Array Cards
- TaqMan Fast Advanced Master Mix
- TaqMan Universal Master Mix II, No UNG
For OpenArray:
TaqMan OpenArray Real-Time PCR Master Mix ›
Run qPCR
Instruments for real time PCR and digital PCR
- QuantStudio 3 Real-Time PCR System—simple to use for all levels of experience
- QuantStudio 5 Real-Time PCR System—easy controls for optimized experiments
- QuantStudio 6 Flex Real-Time PCR System—high flexibility and intuitive design
- QuantStudio 7 Flex Real-Time PCR System—high versatility and intuitive design
- QuantStudio 12k Flex Real-Time PCR System—maximum throughput, flexibility, and scalability
Analyze
Process and interpret qPCR data with intuitive software data analysis tools for a range of applications
Isolate RNA
Isolate RNA using methods that preserve RNA integrity and expression profiles
- RNA isolation kits
Innovative Ambion and Invitrogen RNA extraction products - mirVana miRNA Isolation kits
These kits use a rapid procedure to isolate total RNA, including small RNAs such as miRNA, small interfering RNA (siRNA), and small nuclear RNA (snRNA), from tissues and cells. We can address a small number of targets or handle samples that require high throughput. - TaqMan miRNA ABC Purification Kit
For use with the classic TaqMan MicroRNA assays, obtain high quality miRNA for sensitive and specific miRNA expression analysis from difficult sample types including blood, serum, plasma, FFPE, solid tissues, cultured cells, saliva, or urine
Design and optimize primers
Not required for Applied Biosystems TaqMan Assays. These assays have passed our industry leading bioinformatics design criteria.
Reverse Transcription
Convert your RNA into cDNA, over a wide range of RNA concentrations.
TaqMan Advanced miRNA cDNA Synthesis Kit
Provides miR-Amp Universal Amplification with universal primers and master mix to increase the amount of cDNA for each target with accurate detection as few as 60 copies from as little as 1pg of total RNA or 2 μL of serum/plasma. Pre-Amplification Step for TaqMan Advanced miRNA Assays.
Note that a pre-amplification step is performed after the RT. The miR-Amp master mix and miR-Amp primer mix are included in the TaqMan Advanced miRNA cDNA Synthesis Kit.
TaqMan MicroRNA Reverse Transcription Kit
Classic TaqMan microRNA kit for use with the TaqMan MicroRNA Assays to convert miRNA to cDNA (miRNA specific RT). Best for 1-10 targets. Not compatible with TaqMan Advanced miRNA products.
Megaplex RT Primers (For TAC and OpenArray)
Designed to streamline experimental workflows, Megaplex RT Primers provide a single reaction solution when preparing cDNA for real-time PCR analysis on a TaqMan MicroRNA Array.
Megaplex PreAmp Primers
Pre-amplification and reverse transcription primer for use with the classic TaqMan MicroRNA assay (not for use with TaqMan Advanced MicroRNA products) when sensitivity is of the utmost importance, Megaplex PreAmp Primers significantly enhance the detection of miRNAs expressed at low levels, enabling the generation of a comprehensive expression profile using very little sample—as little as 1 ng of input total RNA.
- MegaPlex PreAmp Primers
- Megaplex Primer Pools, Human Pools Set v3.0
- Megaplex Primer Pools, Rodent Pools Set v3.0
Note: When used with - OpenArray, TAC with less than 350ng of RNA and optional for TaqMan Array Cards with greater than 350ng of RNA. For single tube assays you can run pre-amplification with the appropriate protocol.
TaqMan PreAmp Master Mix
Stretch as little as 1 ng of cDNA into 200 real-time PCR reactions for gene expression analysis using TaqMan Gene Expression assays
Amplify cDNA
- TaqMan Advanced miRNA Assays
Enable highly sensitive and specific quantification of mature miRNAs using qPCR. This streamlined workflow includes universal reverse transcription (RT)—one RT step for all miRNA targets. - TaqMan MicroRNA Assays
A simple, two-step protocol includes reverse transcription with a miRNA-specific primer, followed by real-time PCR with TaqMan assays - TaqMan Array miRNA Cards and Plates
Enable highly sensitive and specific quantification of miRNAs in a high throughput format. Both classic and Advanced chemistry cards and plates are available. - Custom TaqMan Advanced miRNA Array Cards
- Custom TaqMan miRNA Array Cards
- TaqMan Fast Advanced Master Mix
- TaqMan Universal Master Mix II, No UNG
For OpenArray:
TaqMan OpenArray Real-Time PCR Master Mix ›
Run qPCR
Instruments for real time PCR and digital PCR
- QuantStudio 3 Real-Time PCR System—simple to use for all levels of experience
- QuantStudio 5 Real-Time PCR System—easy controls for optimized experiments
- QuantStudio 6 Flex Real-Time PCR System—high flexibility and intuitive design
- QuantStudio 7 Flex Real-Time PCR System—high versatility and intuitive design
- QuantStudio 12k Flex Real-Time PCR System—maximum throughput, flexibility, and scalability
Analyze
Process and interpret qPCR data with intuitive software data analysis tools for a range of applications
Additional miRNA categories
We have more options for performing microRNA and ncRNA studies using real-time PCR.
- TaqMan Non-coding RNA Assays
For reliable detection and quantitation of non-coding human, mouse, or rat transcripts longer than 200 nucleotides - TaqMan Pri-miRNA Assays
Detect and measure primary miRNAs from human, mouse, or rat species - Custom TaqMan Array miRNA Cards
TaqMan™ Array MicroRNA Cards are the ideal solution for profiling large numbers of miRNAs.
- Custom TaqMan Small RNA Assays
Custom assays are ideal to quantitate your small RNA when a predesigned assay is unavailable—all you need is the sequence of interest. - miRNA Mimics & Inhibitors
More than 50,000 miRNA mimics that mimic endogenous miRNAs and enable miRNA functional analysis by up-regulation of miRNA activity and Inhibitors designed to specifically bind to and inhibit endogenous miRNA molecules and enable miRNA functional analysis by down-regulation of miRNA activity.
For Research Use Only. Not for use in diagnostic procedures.