a simple way to evaluate self-designed probes for tumor specific multiplex ligation-dependent probe amplification (mlpa)一个简单的方法来评估自行设计探针对肿瘤特定的多路复用ligation-dependent探测器放大(mlpa).pdfVIP

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a simple way to evaluate self-designed probes for tumor specific multiplex ligation-dependent probe amplification (mlpa)一个简单的方法来评估自行设计探针对肿瘤特定的多路复用ligation-dependent探测器放大(mlpa).pdf

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a simple way to evaluate self-designed probes for tumor specific multiplex ligation-dependent probe amplification (mlpa)一个简单的方法来评估自行设计探针对肿瘤特定的多路复用ligation-dependent探测器放大(mlpa)

Pedersen et al. BMC Research Notes 2010, 3:179 /1756-0500/3/179 T E C H N I C A L N O T E Open Access Technical Note A simple way to evaluate self-designed probes for tumor specific Multiplex Ligation-dependent Probe Amplification (MLPA) †1,2,4 †1,2 3,5 2 1 Kristina Pedersen , Emilia Wiechec , Bo E Madsen , Jens Overgaard and Lise Lotte Hansen* Abstract Background: The Multiplex Ligation-dependent Probe Amplification (MLPA) is widely used for analysis of copy number variations (CNVs) in single or multiple loci. MLPA is a versatile methodology and important tool in cancer research; it provides precise information on increased or decreased copy number at specific loci as opposed to loss of heterozygosity (LOH) studies based upon microsatellite analysis. Pre-designed MLPA kits and software are commercially available to analyze multiple exons, genes, and genomic regions. However, an increasing demand for new gene specific assays makes it necessary to self-design new MLPA probes for which the available software may not be applicable. During evaluation of new self-designed reference probes, we encountered a number of problems, especially when applying the MLPA methodology to tumor samples. Findings: DNA samples from 48 unaffected individuals and 145 breast cancer patients were used to evaluate 11 self- designed MLPA probes and determine the cut-off values for CNV, before applying the MLPA probes to normalize the target probes in a cohort of affected individuals. To test the calculation strategy, three probes were designed to cover regions in Regulator of G-protein Signaling 8 (RGS8), which we previously have identified as being affected by allelic imbalance by LOH analysis across RGS8 in the cohort