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Transcriptome pp Transcriptome pp Document Transcript

  • ProductProfile SureFIND Transcriptome PCR Array For discovering regulators of gene expression The SureFIND Transcriptome PCR Array identifies which miRNA, protein, or pathway regulates the expression of any gene. In each well of a SureFIND Transcriptome PCR Array, is a unique “transcriptome” or cDNA sample synthesized from cells treated with a unique miRNA mimic, siRNA, or chemical inhibitor. A single qPCR reaction screens 90 different regulators. An expression change in a specific well reveals a transcriptional regulator for the tested gene. The SureFIND Transcriptome PCR Array provides: n Rapid discovery of gene expression regulators n Pre-loaded arrays for simple setup n Compatibility with SYBR® Green or hydrolysis probe assays n Protocol overview Included data analysis software Prepare reaction mix with genespecific assay and master mix Load SureFIND Transcriptome PCR Array with reaction mix Greater savings and unparalleled ease of use SureFIND Transcriptome PCR Array layout The SureFIND Transcriptome PCR Array is a single PCR plate with data 01 03 04 05 06 07 08 09 10 11 14 15 16 17 18 19 20 21 22 23 24 25 synthesized from an experimentally treated cell sample. The cells are 02 13 analysis software. Each well (Figure 1) has a unique cDNA sample 12 26 27 28 29 30 31 32 33 34 35 36 treated with a panel of miRNA mimics, chemicals, or siRNAs. By 37 38 39 40 41 42 43 44 45 46 47 48 correlation, a treatment that changes the expression of a gene is identified 49 50 51 52 53 54 55 56 57 58 59 60 as a candidate regulator. Therefore, the SureFIND Transcriptome PCR 61 62 63 64 65 66 67 68 69 70 71 72 79 80 81 82 83 84 73 74 75 76 77 78 85 Arrays allow you to screen 90 different regulators in a single assay. 86 87 88 89 90 VTC VTC VTC VTC PPC PPC To perform the assay, add a gene-specific qPCR assay to every well in Preloaded cDNA samples the array and run it in a qPCR instrument. SYBR Green and probe-based Vehicletreated controls Positive PCR controls qPCR assays are compatible with these arrays. CT values obtained are imported into the software to identify regulators of the selected 1.E+001 calculations and delivers a list of which miRNA, transcription factor, or pathway regulates the tested gene. Delta Rn gene’s expression. The software performs the ΔΔCT based fold-change Thermocycling run in qPCR instrument 1.E-000 1.E-001 1.E-002 1.E-003 4 8 12 16 20 24 28 32 36 40 Cycle number Figure 1. Protocol overview. SureFIND Transcriptome PCR Arrays preloaded with cDNA synthesized from 90 different experimentally treated cell samples. Each unique cDNA sample is derived from cells treated with a miRNA mimic, chemical, or siRNA. After a qPCR reaction for a gene of interest, the software identifies which miRNA, protein, or pathway regulates that gene. Identify transcriptional regulators: Export CT values to data analysis software Sample & Assay Technologies
  • Application experiment Expression fold change (log2) 0.5 0 -0.5 -1 -1.5 8 37 N 3 m iR A-2 N 9 A- b 13 5b 13 m iR A- A- N N iR iR m m AN iR m m iR N A- 19 20 5 3b -2 Figure 2. SureFIND Transcriptome PCR Arrays identify miRNA regulators of INPPL1 expression. PCR reactions using primers for INPPL1 (SHIP2) and a normalization gene were performed using the SureFIND Cancer miRNA Transcriptome PCR Array. The SureFIND Cancer miRNA Transcriptome PCR Array contains cDNA samples derived from HeLa cells treated with 90 different cancer-associated miRNA mimics. The software identified miRNA-205, -193b, -138, -373, -29b, and -135b as negative regulators of INPPL1 expression. Ordering Information Product Contents Cat. no. SureFIND miRNA Transcriptome PCR Array For identifying miRNAs that regulate the expression of any gene with cDNAs derived from HeLa cells treated with miRNA mimics. 336611 SureFIND Pathway Transcriptome PCR Array For identifying pathways that regulate the expression of any gene with cDNAs derived from HEK293, HeLa or MCF-7 cells treated with chemical inhibitors. 336611 SureFIND Transcription Factor Transcriptome PCR Array For identifying transcription factors that regulate the expression of any gene with cDNAs derived from MCF-7 cells treated with siRNAs. 336611 For up-to-date licensing information and product-specific disclaimers, see the respective handbook or user manual. User manuals are available at www.SABiosciences.com or can be requested from QIAGEN Technical Services or your local distributor. These kits are intended for molecular biology applications. Not intended for diagnostic purposes. Trademarks: QIAGEN® (QIAGEN Group); SYBR® (Molecular Probes Inc.). 1066551 11/2010 © 2010 QIAGEN, all rights reserved. www.SABiosciences.com Australia n 1-800-243-066 Austria n 00800-22448000 Belgium n 00800-22448000 Brazil n 0800-557779 Canada n 0800-362-7737 China n 0800-988-0325 Denmark n 00800-22448000 Finland n 00800-22448000 France n 00800-22448000 Germany n 00800-22448000 Ireland n 00800-22448000 Italy n 00800-22448000 Japan n 03-5632-9610 Luxembourg n 00800-22448000 Mexico n 01-800-7742-436 Netherlands n 00800-22448000 Norway n 00800-22448000 Singapore n 1800-742-4368 Sweden n 00800-22448000 Switzerland n 00800-22448000 UK n 00800-22448000 USA n 1-888-503-3187 Sample & Assay Technologies