Moderate degradation does not preclude microarray analysis of small amounts of RNA

Gene expression analysis by microarrays using small amounts of RNA is becoming more and more popular against the background of advances and increasing importance of small-sample acquisition methods like laser microdissection techniques. The quality of RNA preparations from such samples constitutes a...

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Main Authors: Oliver Schoor, Toni Weinschenk, Jörg Hennenlotter, Stefan Corvin, Arnulf Stenzl, Hans-Georg Rammensee, Stefan Stevanović
Format: Article
Language:English
Published: Taylor & Francis Group 2003-12-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/03356rr01
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author Oliver Schoor
Toni Weinschenk
Jörg Hennenlotter
Stefan Corvin
Arnulf Stenzl
Hans-Georg Rammensee
Stefan Stevanović
author_facet Oliver Schoor
Toni Weinschenk
Jörg Hennenlotter
Stefan Corvin
Arnulf Stenzl
Hans-Georg Rammensee
Stefan Stevanović
author_sort Oliver Schoor
collection DOAJ
description Gene expression analysis by microarrays using small amounts of RNA is becoming more and more popular against the background of advances and increasing importance of small-sample acquisition methods like laser microdissection techniques. The quality of RNA preparations from such samples constitutes a frequent issue in this context. The aim of this study was to assess the impact of different extents of RNA degradation on the expression profile of the samples. We induced RNA degradation in human tumor and healthy tissue samples by endogeneous ribonucleases. Next, we amplified 20 ng total RNA degraded to different extents by two rounds of in vitro transcription and analyzed them using Affymetrix oligonucleotide microarrays. Expression differences for some genes were independently confirmed by real-time quantitative PCR. Our results suggest that gene expression profiles obtained from partially degraded RNA samples with still visible ribosomal bands exhibit a high degree of similarity compared to intact samples and that RNA samples of suboptimal quality might therefore still lead to meaningful results if used carefully.
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spelling doaj-art-41899b51eee44635967b71fe552dbfe72025-08-20T02:26:00ZengTaylor & Francis GroupBioTechniques0736-62051940-98182003-12-013561192120110.2144/03356rr01Moderate degradation does not preclude microarray analysis of small amounts of RNAOliver Schoor0Toni Weinschenk1Jörg Hennenlotter2Stefan Corvin3Arnulf Stenzl4Hans-Georg Rammensee5Stefan Stevanović61University of Tübingen, Tübingen, Germany1University of Tübingen, Tübingen, Germany1University of Tübingen, Tübingen, Germany1University of Tübingen, Tübingen, Germany1University of Tübingen, Tübingen, Germany1University of Tübingen, Tübingen, Germany1University of Tübingen, Tübingen, GermanyGene expression analysis by microarrays using small amounts of RNA is becoming more and more popular against the background of advances and increasing importance of small-sample acquisition methods like laser microdissection techniques. The quality of RNA preparations from such samples constitutes a frequent issue in this context. The aim of this study was to assess the impact of different extents of RNA degradation on the expression profile of the samples. We induced RNA degradation in human tumor and healthy tissue samples by endogeneous ribonucleases. Next, we amplified 20 ng total RNA degraded to different extents by two rounds of in vitro transcription and analyzed them using Affymetrix oligonucleotide microarrays. Expression differences for some genes were independently confirmed by real-time quantitative PCR. Our results suggest that gene expression profiles obtained from partially degraded RNA samples with still visible ribosomal bands exhibit a high degree of similarity compared to intact samples and that RNA samples of suboptimal quality might therefore still lead to meaningful results if used carefully.https://www.future-science.com/doi/10.2144/03356rr01
spellingShingle Oliver Schoor
Toni Weinschenk
Jörg Hennenlotter
Stefan Corvin
Arnulf Stenzl
Hans-Georg Rammensee
Stefan Stevanović
Moderate degradation does not preclude microarray analysis of small amounts of RNA
BioTechniques
title Moderate degradation does not preclude microarray analysis of small amounts of RNA
title_full Moderate degradation does not preclude microarray analysis of small amounts of RNA
title_fullStr Moderate degradation does not preclude microarray analysis of small amounts of RNA
title_full_unstemmed Moderate degradation does not preclude microarray analysis of small amounts of RNA
title_short Moderate degradation does not preclude microarray analysis of small amounts of RNA
title_sort moderate degradation does not preclude microarray analysis of small amounts of rna
url https://www.future-science.com/doi/10.2144/03356rr01
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