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TissueScan Cancer qPCR Array Cancer Biomarker Validation Using Archived Tumor Tissues
Sponsored,vendor-submitted protocol   Sponsored by Origene Technologies    Published in BioTechniques Protocol Guide 2009 (p.13) DOI: 10.2144/000113033 Abstract

TissueScan is a unique qPCR array for fast and accurate gene expression profiling across a large number of cancer patient samples. High-quality cDNAs were prepared from thousands of well-documented cancer biopsy samples, normalized, and assembled into ready-to-use gene expression panels. This alleviates for researchers the tedious work of large sample collection and the meticulous work of sample preparation, while facilitating quick and reliable profiling of gene expression levels across cancer progression stages. TissueScan combines the high sensitivity and specificity of the RT-PCR methodology with a well-designed multi-sample format. It is an excellent tool for validation of potential cancer markers such as those obtained by microarray or differential display.

TissueScan is a unique qPCR array for fast and accurate gene expression profiling across a large number of cancer patient samples. High-quality cDNAs were prepared from thousands of well-documented cancer biopsy samples, normalized, and assembled into ready-to-use gene expression panels. This alleviates for researchers the tedious work of large sample collection and the meticulous work of sample preparation, while facilitating quick and reliable profiling of gene expression levels across cancer progression stages. TissueScan combines the high sensitivity and specificity of the RT-PCR methodology with a well-designed multi-sample format. It is an excellent tool for validation of potential cancer markers such as those obtained by microarray or differential display.

Application

To illustrate the validity of TissueScan, we used the Lung Cancer Panel (HLRT-101) to evaluate the expression level of topoisomerase II-alpha (TOPO II), which has been known to be upregulated in a large percentage of lung cancers. A pair of primers was designed to detect the transcript of this gene. SYBR Green I based real-time PCR was performed. The raw data (Figure 1) and the expression level plot calculated by the Ct method (Figure 2) are shown below. A second panel in the kit was used with beta-actin control primers provided in the kit (Figure 3).


Figure 1. Real-time PCR results with TOPO II primers. (Click to enlarge)



Figure 2. Normalized relative expression of TOPO II alpha among 48 lung samples. (Click to enlarge)



Figure 3. Real-time PCR results with beta-actin primers. (Click to enlarge)


Results

The results showed that TOPO II expression levels are markedly higher than in the normal tissues, supporting the notion that topoisomerase can serve as a valid biomarker for lung cancer. Products

The TissueScan kit includes two identical 48-well panels, beta-actin control primers, and a copy of the Application Guide. The normalized cDNAs are lyophilized in 48-well PCR plates. A sample list is shown in Table 1. The most up-to-date list can be found on our web site at www.origene.com/geneexpression.


Table 1.


Correspondence

Address correspondence to OriGene, Your Gene Company, 6 Taft Court, Rockville, MD 20850, USA; Tel.: 1-888-267-4436; [email protected]; www.origene.com.

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