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BMC Genomics

, 7:34

First Online: 24 February 2006Received: 12 July 2005Accepted: 24 February 2006

Abstract

BackgroundLow density arrays LDAs have recently been introduced as a novel approach to gene expression profiling. Based on real time quantitative RT-PCR QRT-PCR, these arrays enable a more focused and sensitive approach to the study of gene expression than gene chips, while offering higher throughput than more established approaches to QRT-PCR. We have now evaluated LDAs as a means of determining the expression of multiple genes simultaneously in human tissues and cells.

ResultsComparisons between LDAs reveal low variability, with correlation coefficients close to 1. By performing 2-fold and 10-fold serial dilutions of cDNA samples in the LDAs we determined a clear linear relationship between the gene expression data points over 5 orders of magnitude. We also showed that it is possible to use LDAs to accurately and quantitatively detect 2-fold changes in target copy number as well as measuring genes that are expressed with low and high copy numbers in the range of 1 × 10 – 1 × 10 copies. Furthermore, the data generated by the LDA from a cell based pharmacological study were comparable to data generated by conventional QRT-PCR.

ConclusionLDAs represent a valuable new approach for sensitive and quantitative gene expression profiling.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2164-7-34 contains supplementary material, which is available to authorized users.

Andrew B Goulter, Daniel W Harmer contributed equally to this work.

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Autor: Andrew B Goulter - Daniel W Harmer - Kenneth L Clark

Fuente: https://link.springer.com/article/10.1186/1471-2164-7-34







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