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A high-density quantitative nuclease protection microarray platform for high throughput analysis of gene expression

Paper ID Volume ID Publish Year Pages File Format Full-Text
23767 43471 2011 8 PDF Available
Title
A high-density quantitative nuclease protection microarray platform for high throughput analysis of gene expression
Abstract

The quantitative nuclease protection assay (qNPA) is a very simple and highly sensitive method for measuring mRNA transcripts, can be used on a variety of sample types, and is amenable to high-throughput sample processing. We have combined the power of the qNPA assay with the density of a DNA microarray to create a qNPA Microarray platform. This platform is compatible with common laboratory equipment: it uses fluorescence-based detection, can be analyzed with common microarray scanners, and is in an SBS footprint with 96-well layout for high-throughput applications. Here, we demonstrate the characteristics of a qNPA Microarray slide that contains up to 1700 gene elements per well. We show that the new platform can reliably detect transcripts at levels as low as 10 fM with median CVs below 12%. On a standardized set of samples, the qNPA Microarray detected the same trends in gene expression as the original qNPA technology, real time qPCR, and Affymetrix GeneChip DNA Microarrays. Given its ease of use, compatibility with multiple sample types, high-throughput capabilities, and its integration with standard laboratory equipment, the qNPA Microarray is a powerful new platform for gene expression research.

Keywords
AO, anchor oligonucleotide; bp, base pair; CV, coefficient of variation; DL, detection linker; DP, detection probe; HTG, High Throughput Genomics, Inc.; NPP, nuclease protection probe; PL, programming linker; qNPA, quantitative nuclease protection assay;
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A high-density quantitative nuclease protection microarray platform for high throughput analysis of gene expression
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 154, Issue 1, 10 June 2011, Pages 68–75
Authors
, , , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us