Comparison of target labeling methods for use with Affymetrix GeneChips
Open Access
- 18 May 2007
- journal article
- research article
- Published by Springer Nature in BMC Biotechnology
- Vol. 7 (1) , 24
Abstract
Background: Several different commercial one-cycle labeling kits are available for preparation of the target for use with the Affymetrix GeneChip platform. However, there have been no evaluations of these different kits to determine if comparable results were generated. We report on the cRNA target synthesis, labeling efficiency and hybridization results using the One-Cycle Target Labeling Assay™ (Affymetrix), the BioArray RNA Amplification and Labeling System™ (Enzo Life Sciences), and the Superscript RNA Amplification System (Invitrogen Life Technologies). Results: The only notable difference between kits was in the yield of cRNA target synthesized during in vitro transcription, where the BioArray assay had to be repeated several times in order to have sufficient target. However, each kit resulted in comparable signal and detection calls when hybridized to the Affymetrix GeneChip. Conclusion: These 3 one-cycle labeling kits produce comparable hybridization results. This provides users with several kit options and flexibility when using the Affymetrix system.Keywords
This publication has 9 references indexed in Scilit:
- Comparative analysis of a 3′ end tag PCR and a linear RNA amplification approach for microarray analysisJournal of Biotechnology, 2007
- In Vitro Transcription Amplification and Labeling Methods Contribute to the Variability of Gene Expression Profiling with DNA MicroarraysThe Journal of Molecular Diagnostics, 2006
- Analysis of Affymetrix GeneChip® Data Using Amplified RNABioTechniques, 2006
- Precision Profiling and Components of Variability Analysis for Affymetrix Microarray Assays Run in a Clinical ContextThe Journal of Molecular Diagnostics, 2005
- RNA amplification for successful gene profiling analysisJournal of Translational Medicine, 2005
- Amplification protocols introduce systematic but reproducible errors into gene expression studiesBioTechniques, 2004
- A compendium of gene expression in normal human tissuesPhysiological Genomics, 2001
- Protein Chips Based on Recombinant Antibody Fragments: A Highly Sensitive Approach as Detected by Mass SpectrometryBioTechniques, 2001
- Amplified RNA synthesized from limited quantities of heterogeneous cDNA.Proceedings of the National Academy of Sciences, 1990