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Pre-miRNA Analysis Using Real-Time PCR
Workflow features
- Ideal for quantifying precursor miRNA or “pre-miRNA”
- Enables the study of miRNA biogenesis when used in combination with Applied Biosystems TaqMan Pri-miRNA Assays and TaqMan MicroRNA Assays
How it works
MicroRNA “genes” are transcribed in the nucleus creating a long pri-miRNA transcript. The pri-miRNA sequence contains one or more stem-loop sequences. These pri-miRNAs are cleaved in the nucleus by Drosha to generate pre-miRNAs, which are then exported into the cytoplasm by Exportin-5, where they are further cleaved by Dicer to produce mature miRNA sequences.
TaqMan Pre-miRNA Assays are designed using Applied Biosystems state-of-the-art bioinformatics pipeline and are designed to be specific against individual stem-loop sequences identified in the Sanger miRBase sequence repository. This helps ensure that the precursor miRNA is accurately measured.
Pre-miRNA analysis experimental workflow
Applied Biosystems offers a variety of kits that preserve small RNAs such as miRNAs. While it is not essential to use them due to the larger size of primary miRNA transcripts, these kits do enable both pre-miRNA and mature miRNA quantitation from the same RNA sample. Each kit is ideal for miRNA analysis because it enables quantitative recovery of all RNA >10 nt in length and maintenance of representative amounts of all RNA (eliminating experimental bias). To enable accurate quantitation with TaqMan Pre-miRNA Assays, samples must be free from genomic DNA.
Recommended products
mirVana miRNA Isolation Kit
Isolate total RNA that contains small RNAs such as miRNA, siRNA, and snRNA. These small RNAs are often lost during traditional RNA isolation methods. In addition, this kit includes reagents and instructions for the isolation of RNA that is enriched for small RNA (<200 bp), enabling more sensitive detection of low-level small RNAs.
The High Capacity cDNA Reverse Transcription Kit provides all the necessary components to reverse transcribe long RNA into cDNA.
Recommended products
Recommended products
TaqMan Pre-miRNA Assays
TaqMan Pre-miRNA Assays quantitate precursor microRNA (pre-miRNA) transcripts, the intermediate RNA molecules in the miRNA biogenesis pathway. TaqMan Pre-miRNA Assays are designed using our state-of-the-art bioinformatics pipeline, delivering gold-standard assay performance and data quality for pre-miRNA studies.
Learn more ›
TaqMan Fast Advanced Master Mix
TaqMan Fast Advanced Master Mix has been designed to match or exceed the performance of standard master mixes, delivering shorter run times (<40 minutes) with results equal to or better than what is achieved today.
Learn more ›
Applied Biosystems real-time PCR instruments
As a leader in PCR innovation, we offer you the gold standard in real-time PCR instrumentation. With our Applied Biosystems real-time PCR platforms, you get true value with excellent performance, reliability, and world-class support. Designed for compatibility with Applied Biosystems TaqMan Assays and the flexibility of unlimited real-time chemistry choices, we make it easy for you to get started with your experiments.
Learn more ›
Analyze your qPCR data using our many different analysis modules.
- Connect cloud-based platform—free to sign up
- Desktop software—downloads can be found through the Support Center for your specific Applied Biosystems qPCR instrument
- ExpressionSuite Software—free to download
Applied Biosystems offers a variety of kits that preserve small RNAs such as miRNAs. While it is not essential to use them due to the larger size of primary miRNA transcripts, these kits do enable both pre-miRNA and mature miRNA quantitation from the same RNA sample. Each kit is ideal for miRNA analysis because it enables quantitative recovery of all RNA >10 nt in length and maintenance of representative amounts of all RNA (eliminating experimental bias). To enable accurate quantitation with TaqMan Pre-miRNA Assays, samples must be free from genomic DNA.
Recommended products
mirVana miRNA Isolation Kit
Isolate total RNA that contains small RNAs such as miRNA, siRNA, and snRNA. These small RNAs are often lost during traditional RNA isolation methods. In addition, this kit includes reagents and instructions for the isolation of RNA that is enriched for small RNA (<200 bp), enabling more sensitive detection of low-level small RNAs.
The High Capacity cDNA Reverse Transcription Kit provides all the necessary components to reverse transcribe long RNA into cDNA.
Recommended products
Recommended products
TaqMan Pre-miRNA Assays
TaqMan Pre-miRNA Assays quantitate precursor microRNA (pre-miRNA) transcripts, the intermediate RNA molecules in the miRNA biogenesis pathway. TaqMan Pre-miRNA Assays are designed using our state-of-the-art bioinformatics pipeline, delivering gold-standard assay performance and data quality for pre-miRNA studies.
Learn more ›
TaqMan Fast Advanced Master Mix
TaqMan Fast Advanced Master Mix has been designed to match or exceed the performance of standard master mixes, delivering shorter run times (<40 minutes) with results equal to or better than what is achieved today.
Learn more ›
Applied Biosystems real-time PCR instruments
As a leader in PCR innovation, we offer you the gold standard in real-time PCR instrumentation. With our Applied Biosystems real-time PCR platforms, you get true value with excellent performance, reliability, and world-class support. Designed for compatibility with Applied Biosystems TaqMan Assays and the flexibility of unlimited real-time chemistry choices, we make it easy for you to get started with your experiments.
Learn more ›
Analyze your qPCR data using our many different analysis modules.
- Connect cloud-based platform—free to sign up
- Desktop software—downloads can be found through the Support Center for your specific Applied Biosystems qPCR instrument
- ExpressionSuite Software—free to download
For Research Use Only. Not for use in diagnostic procedures.