使用实时荧光定量PCR进行长链非编码RNA (ncRNA) 分析

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

它如何工作

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

Step 1Isolate RNA
Step 2Reverse Transcribe RNA into cDNA
Step 3Amplify cDNA (optional)
Step 4Set Up & Run qPCR Reactions
Step 5Analyze Data

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.

推荐产品

		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. mirVana™ miRNA Isolation Kit mirVana™ miRNA Isolation Kit (without Phenol)
		RecoverAll™ 针对FFPE的总核酸分离试剂盒 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. RecoverAll™ 针对FFPE的总核酸分离试剂盒
		*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. *TURBO DNA-free™* *TURBO DNA-free™ w/Manual*
		TaqMan® microRNA Cells-to-CT™试剂盒 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. TaqMan® microRNA Cells-to-CT™试剂盒 TaqMan® MicroRNA Cells-to-CT™ Kit w/Manual Cells-to-CT Bulk Lysis Reagents Cells-to-CT™ Stop Solution

Step 2 : Reverse Transcribe RNA into cDNA

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

文献

应用说明

TaqMan Non-coding RNA Assays Help Characterize Hidden RNA Regulatory Networks

产品信息

TaqMan Non-coding RNA Assays (Product Bulletin)

仅用于研究用途。不得用于任何动物或人类的治疗或诊断。

		TaqMan®非编码RNA分析 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®通用预混液II TaqMan® Universal Master Mix II offers convenience and reliability for all real-time applications including microRNA, gene expression, and genotyping studies.

		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.