An mi script-serum
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This document describes a study that used miRNA expression profiling to identify potential biomarkers for lung cancer. Researchers used the miRNeasy Serum/Plasma Kit to isolate total RNA from normal and lung cancer serum samples. They then used the miScript PCR System and the Human Serum & Plasma 384HC miScript miRNA PCR Array to analyze miRNA expression levels. Several miRNAs were found to be significantly upregulated in lung cancer serum samples, including miRNAs that have previously been identified as blood-based markers for lung cancer.
An mi script-serum
- 1. ApplicationNote Discover miRNA biomarkers in serum or plasma using miScript® miRNA PCR Arrays Abigail Harris, Marcus Lewis, and Jonathan M. Shaffer, Ph.D. Changes in miRNA expression levels are indicative of a number of biological and disease states, and the stability of miRNAs in serum and plasma makes them attractive noninvasive biomarker candidates for cancer and other diseases. Here, we use the miRNeasy Serum/Plasma Kit in conjunction with the miScript PCR System in a pilot experiment to identify potential biomarkers for lung cancer. 1.E+01 QIAGEN provides a sample-to-result solution for miRNA expression 1.E+00 profiling using serum and plasma samples (Figure 2). Total RNA is isolated using the miRNeasy Serum/Plasma Kit in conjunction with the miRNeasy Serum/Plasma Spike-In Control. miRNA expression is then quantified using the miScript PCR System which consists of the miScript II RT Kit, miScript SYBR® Green PCR Kit, and miScript miRNA PCR Arrays. miScript miRNA PCR Arrays contain miRNA-specific assays (miScript 1.E-03 T 1.E-04 1.E+01 1.E+00 1.E-01 1.E-02 In addition to facilitating screening of the whole miRNome, specific 1.E-02 1.E-03 using a state-of-the-art bioinformatics algorithm and text mining tools. 1.E-01 1.E-04 Primer Assays) organized into biological and disease-focused panels Lung cancer serum (2-∆C ) Discover biomarkers for cancer, diabetes, and more Normal serum (2-∆C ) T solutions have been developed for miRNA expression profiling using RNA from serum or plasma samples (Table 1). In this example, the Human Serum & Plasma 384HC miScript miRNA PCR Array has been used to profile miRNA expression changes in total RNA from lung cancer serum (Figure 1). Table 1. miScript miRNA PCR Arrays for profiling miRNA expression in serum or plasma samples miScript miRNA PCR Array Use miRNome Profile the expression of the entire collection of available miScript Primer Assays Serum & Plasma Profile the expression of the 84 miRNAs most commonly reported to be differentially upregulated in serum or plasma samples in cancer and other diseases Serum & Plasma 384HC Figure 1. miRNA expression changes identified in serum from lung cancer donors using Human Serum & Plasma 384HC miScript miRNA PCR Array. Total RNA was isolated from 200 µl normal and lung cancer serum samples (3 each) using the miRNeasy Serum/Plasma Kit. The Human Serum & Plasma 384HC miScript miRNA PCR Array, in combination with the miScript PCR System, was used to profile mature miRNA expression by real-time PCR. The miRNeasy Serum/Plasma Spike-In Control was used for normalization. In total, the expression of 183 mature miRNAs was detected. Included in the miRNAs that exhibited significant upregulation are hsa-miR-25, hsa-miR-223, and hsa-miR-17*, which have previously been shown to be blood-based markers for patients with lung cancer (1). Profile the expression of the 372 miRNAs with detectable expression in serum and plasma samples that are profiled with the miScript PCR System Sample & Assay Technologies
- 2. Figure 2. QIAGEN workflow for miRNA profiling in serum and plasma samples. Ordering Information Product Contents Cat. no. miRNome miScript miRNA PCR Array Full miRNome coverage Varies miScript miRNA PCR Array Pathway- or disease-focused panels of miRNA assays Varies Custom miScript miRNA PCR Array Custom panels of miRNA assays Varies miRNeasy Serum/Plasma Kit (50) Columns, plasticware, and reagents for 50 preps 217184 miRNeasy Serum/Plasma Spike-In Control 10 pmol lyophilized C. elegans miR-39 miRNA mimic 219610 miScript SYBR Green PCR Kit (200) Reagents for 200 x 50 μl PCR reactions 218073 miScript Primer Assay (100) miRNA-specific primer for 100 x 50 μl PCR reactions Varies* miScript II RT Kit (12) Reagents for 12 x 20 μl cDNA synthesis reactions 218160 * Visit GeneGlobe to browse and order these products (www.qiagen.com/GeneGlobe). Discover more at www.qiagen.com/miRNA For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at www.qiagen.com or can be requested from QIAGEN Technical Services or your local distributor. Reference 1. Chen X, et al. (2008) Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res. 18, 997. Trademarks: QIAGEN®, miScript® (QIAGEN Group); SYBR® (Life Technologies) 1071213 05/2012 © 2012 QIAGEN, all rights reserved. Australia n 1-800-243-800 Austria n 0800-281011 Belgium n 0800-79612 Brazil n 0800-557779 Canada n 800-572-9613 China n 800-988-0325 Denmark n 80-885945 Finland n 0800-914416 France n 01-60-920-930 Germany n 02103-29-12000 Hong Kong n 800 933 965 India n 1-800-102-4114 Ireland n 1800 555 049 Italy n 800-787980 Japan n 03-6890-7300 Korea (South) n 080-000-7145 Luxembourg n 8002 2076 Mexico n 01-800-7742-436 The Netherlands n 0800-0229592 Norway n 800-18859 Singapore n 1800-742-4368 Spain n 91-630-7050 Sweden n 020-790282 Switzerland n 055-254-22-11 Taiwan n 0080-665-1947 UK n 01293-422-911 USA n 800-426-8157 www.qiagen.com www.SABiosciences.com Sample & Assay Technologies