Effect of target length on specificity and sensitivity of oligonucleotide microarrays: A comparison between dendrimer and modified PCR based labelling methods

SENSORS AND ACTUATORS B-CHEMICAL

卷:198

Article

Gibriel, Abdullah A. Y.^1,2 

[1] Ahram Canadian Univ, Fac Pharm, Dept Biochem & Mol Biol, 6th Of October 11728, Egypt

[2] Univ Glasgow, Coll Med Vet & Life Sci, Inst Mol Cell & Syst Biol, Glasgow, Lanark, Scotland

关键词: Oligonucleotide microarrays;    Secondary structure;    Hybridization specificity;    Sensitivity;    PCR;    Dendrimer;   

DOI  :  10.1016/j.snb.2014.03.038

来源: Elsevier

【 摘 要 】

DNA microarrays are widely used as end point detectors for gene expression analysis. Several methods have been developed for target labelling to enable quantification but without taking target length into consideration. Here we highlight the importance of choosing the optimum target length that would ensure specificity without compromising sensitivity of the assay. For this, eight plasmids that are identical to each other except for a closely related 23 bp unique reporter (UR) sequence were used to examine the hybridization efficiency for these URs. Targets of various lengths were generated and labelled as follows: full length and 330 bases transcripts using a dendrimer labelling method, 120 bp amplicons by the modified PCR end labelling method and synthetic labelled targets of 33 bases. This report also shows the advantages of using the modified PCR method over other labelling methods in generating labelled amplicons of the desired lengths to maximize hybridization efficiency. (C) 2014 Elsevier B.V. All rights reserved.

【 授权许可】

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