Workflow Features

  • Ideal for reliably quantitating transcripts longer than 200 nucleotides
  • Detect only the intended noncoding RNA transcript
  • Assays are based on proven TaqMan® Assay technology and use existing assay design and manufacturing infrastructure
  • Available for human, mouse, and rat species

See Literature/References

How It Works

Long noncoding RNAs (ncRNAs) are transcripts longer than 200 nucleotides. This definition distinguishes long ncRNAs from small regulatory RNAs such as miRNAs. Rather than potentially detecting different transcript variants at a particular gene locus like TaqMan® Gene Expression Assays, TaqMan® Non-coding RNA Assays detect only the intended ncRNA transcript. Assays were designed using transcripts from a number of different sources, including NCBI RefSeq NR, RNAdb, and from the content on Invitrogen™ NCode™ Non-coding RNA Arrays.

Long noncoding RNA (ncRNA) Analysis Experimental Workflow

Life Technologies offers a variety of kits that preserve small RNAs. Each kit is ideal for use in long noncoding RNA analysis because they enable quantitative recovery of all RNA > 10nt in length and maintenance of representative amounts of all RNA (eliminating experimental bias). To ensure accurate quantitation with TaqMan® Non-coding RNA 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.

 

RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE
Extract total RNA or DNA from formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues. This kit enables the molecular analysis of archived tissue samples at both the genomic and gene expression levels. With this kit, nucleic acid is obtained that is suitable for RT-qPCR, mutation screening, and microarray analyses.

 

TURBO DNA-free™ Kit
TURBO™ DNase is a recombinant, engineered form of DNase I that is more efficient than wild-type DNase I at digesting trace amounts of unwanted DNA in RNA samples. The TURBO DNA-free™ Kit comes with all reagents needed to perform 50 TURBO™ DNase treatments plus a highly effective DNase Inactivation Reagent, making this kit the tool of choice for RT-PCR sample preparation.

 

TaqMan® MicroRNA Cells-to-CT™ Kit
Featuring a unique method for lysing cultured cells while removing genomic DNA and preserving RNA integrity, the TaqMan® MicroRNA Cells-to-CT™ Kit makes it possible to perform miRNA expression profiling directly from cultured cells without RNA purification.

The High Capacity RNA-to-cDNA kit provides all the necessary components to reverse transcribe long RNA into cDNA. (Note: this step can be omitted if a one-step RT-PCR protocol is used.)

Recommended Products

 

High Capacity RNA-to-cDNA Kit
The High Capacity RNA-to-cDNA™ Kit provides high quality reverse transcription that delivers a streamlined workflow with maximum sensitivity and dynamic range.

TaqMan® PreAmp Master Mix preamplifies small amounts of cDNA without introducing amplification bias to the sample. Preamplification enables you to stretch your precious sample into many more real-time PCR reactions.

Recommended Products

 

TaqMan® PreAmp Master Mix
TaqMan® PreAmp Master Mix preamplifies small amounts of cDNA without introducing amplification bias to the sample. Preamplification enables you to stretch your precious sample into many more real-time PCR reactions.

For the final quantitation step, the cDNA product generated in step 2 is mixed with TaqMan® Gene Expression Master Mix and added to the matched TaqMan® Non-coding RNA Assay.

Recommended Products

 

TaqMan® Non-coding RNA Assays
TaqMan® Non-coding RNA Assays provide a highly reliable way to measure expression of long non-coding transcripts. These assays are based on the same gold standard TaqMan® Assays technology that is used in the TaqMan® Gene Expression Assays collection. With TaqMan® Assay technology, you get the high specificity and sensitivity you need for quantification and validation studies.

 

TaqMan® Universal Master Mix II
TaqMan® Universal Master Mix II offers convenience and reliability for all real-time applications including microRNA, gene expression, and genotyping studies.

Integromics RealTime StatMiner® Software is a powerful, easy-to-use bioinformatics tool for quality control and differential expression analysis of real-time PCR data. The software easily imports raw data or Ct values from any Applied Biosystems® Real-Time PCR System and guides you though every step of analysis for accurate results in minutes.

Recommended Products

 

RealTime StatMiner® Software (optional)
RealTime StatMiner® Software from Integromics is a powerful, easy-to-use bioinformatics tool for quality control and differential expression analysis of real-time PCR data. With just a single click, the software imports raw data or Ct values from any of our real-time PCR instruments, and then guides you through every step of analysis, giving you accurate results in minutes.

 DataAssist™ Software (optional)
DataAssist™ Software is an easy to use data analysis tool that utilizes the comparative Ct (ddCt) method to rapidly and accurately quantitate relative gene expression across a large number of genes and samples.

Literature

For research use only. Not intended for any animal or human therapeutic or diagnostic use.