title: RT2 miRNA Mouse Genome PCR Array
status: Public on Feb 05 2010
contact.city: Jefferson
contact.first: Zhiguang Li
contact.email: zhiguangli88@gmail.com, tao.chen@fda.hhs.gov
contact.state: AR
contact.address: HFT130, 3900 NCTR Road
contact.department: Division of Genetic an Reproductive Toxicology
contact.country: USA
contact.institute: National Center for Toxicological Research
contact.phone: 870-543-7295 870-543-7954
organism: Mus musculus
manufacture_protocol: MicroRNA (miRNA), first discovered in C. elegans in early 1990s, are a class of naturally occurring small RNA molecules, generally 17 to 30 nucleotides long, processed from much larger stem-loop structures (pri-miRNA and pre-miRNA). Mature miRNA is recognized by and guides the RNA-Induced Silencing Complex (RISC) to specific messenger RNA transcript sequences, causing either translation repression or mRNA degradation. This phenomenon adds even more complexity to the regulation of gene expression. Each miRNA can regulate multiple mRNA targets, and a single target mRNA may be regulated by multiple miRNA sequences. Researchers are currently correlating miRNA expression profiles to biological phenotypes in attempts to better understand miRNA-based regulation of gene or mRNA expression.
  The RT2 miRNA PCR Array, a patent-pending technology from SABiosciences, is the latest innovation designed to analyze miRNA expression using real-time, reverse transcription PCR, or qRT-PCR, the most sensitive and reliable method for nucleic acid expression analysis. The arrays take advantage of a SYBR Green real-time PCR detection system optimized for high performance to analyze the expression of many mature miRNA sequences simultaneously. Each 96- or 384-well array plate contains a panel of primer sets for a thoroughly researched set of 88 or 376 relevant, pathway- or disease-focused miRNA, plus four housekeeping assays and two RNA and PCR quality controls. The RT2 Genome miRNA PCR Arrays reflect miRNA sequences annotated by the Sanger miRBase Release 14, and are available for the human (704/752:96/384), mouse (528/579:96/384), and rat (352/370:96/384). (See Figure 2 for the layout of a typical PCR Array.) The specificity and single mismatch discrimination, sensitivity and dynamic range, and reproducibility of the miRNA expression analysis is guaranteed when using the complete miRNA PCR Array System.
  To complete the miRNA PCR Array procedure, reverse transcribe your experimental small RNA samples into first strand cDNA, the template for the PCR, using our RT2 miRNA First Strand Kit. (See Figure 1 for an overview of the miRNA PCR Array procedure.) Then, mix the template with one of our instrument-specific and ready-to-use RT2 SYBR Green qPCR Master Mixes. Aliquot the mixture into each well of the same plate containing the predispensed miRNA-specific assays. Perform PCR, and finally determine relative expression with your real-time instrument and the Ct method. The simplicity of the miRNA PCR Arrays makes them accessible for routine use in every research laboratory.
  Platform instruction link: http://www.sabiosciences.com/Manual/mirnapcrarray.pdf