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Performance of different small sample RNA amplification techniques for hybridization on Affymetrix GeneChips

Paper ID Volume ID Publish Year Pages File Format Full-Text
25361 43570 2007 7 PDF Available
Title
Performance of different small sample RNA amplification techniques for hybridization on Affymetrix GeneChips
Abstract

A key issue in RNA amplification techniques is the preservation of original transcript abundance, however popular high-grade RNA amplification methods lack sufficient validation regarding the potential bias of gene expression profiles. This study evaluated a double-round T7-based and a PCR-based amplification protocol, using the Affymetrix GeneChip platform. Both small sample methods performed excellently in terms of yield and reproducibility (r > 0.99), and also the within-method concordance with respect to differential gene expression was as high as with standard single-round T7-based amplification. However, when comparing the overlap of all differentially expressed genes between standard and small sample methods, this was only moderate for the double-round T7 (48.7–55.0%) as well as for the PCR-based amplification protocol (51.9–58.0%). In contrast, the concordance for the top 100 genes with highest fold changes was significantly higher, indicating that both small sample methods generate reliable results when focusing on strongly regulated genes.

Keywords
DNA microarrays; RNA amplification techniques; Methodology; Evaluation; Concordance
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Performance of different small sample RNA amplification techniques for hybridization on Affymetrix GeneChips
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 129, Issue 4, 10 May 2007, Pages 628–634
Authors
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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