Deep-sequencing protocols influence the results obtained in small-RNA sequencing.
Second-generation sequencing is a powerful method for identifying and quantifying small-RNA components of cells. However, little attention has been paid to the effects of the choice of sequencing platform and library preparation protocol on the results obtained. We present a thorough comparison of s...
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Public Library of Science (PLoS)
2012
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oai:doaj.org-article:1b959c8571564678af790e407e5721812021-11-18T07:26:36ZDeep-sequencing protocols influence the results obtained in small-RNA sequencing.1932-620310.1371/journal.pone.0032724https://doaj.org/article/1b959c8571564678af790e407e5721812012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22384282/?tool=EBIhttps://doaj.org/toc/1932-6203Second-generation sequencing is a powerful method for identifying and quantifying small-RNA components of cells. However, little attention has been paid to the effects of the choice of sequencing platform and library preparation protocol on the results obtained. We present a thorough comparison of small-RNA sequencing libraries generated from the same embryonic stem cell lines, using different sequencing platforms, which represent the three major second-generation sequencing technologies, and protocols. We have analysed and compared the expression of microRNAs, as well as populations of small RNAs derived from repetitive elements. Despite the fact that different libraries display a good correlation between sequencing platforms, qualitative and quantitative variations in the results were found, depending on the protocol used. Thus, when comparing libraries from different biological samples, it is strongly recommended to use the same sequencing platform and protocol in order to ensure the biological relevance of the comparisons.Joern ToedlingNicolas ServantConstance CiaudoLaurent FarinelliOlivier VoinnetEdith HeardEmmanuel BarillotPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 2, p e32724 (2012) |
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Medicine R Science Q Joern Toedling Nicolas Servant Constance Ciaudo Laurent Farinelli Olivier Voinnet Edith Heard Emmanuel Barillot Deep-sequencing protocols influence the results obtained in small-RNA sequencing. |
description |
Second-generation sequencing is a powerful method for identifying and quantifying small-RNA components of cells. However, little attention has been paid to the effects of the choice of sequencing platform and library preparation protocol on the results obtained. We present a thorough comparison of small-RNA sequencing libraries generated from the same embryonic stem cell lines, using different sequencing platforms, which represent the three major second-generation sequencing technologies, and protocols. We have analysed and compared the expression of microRNAs, as well as populations of small RNAs derived from repetitive elements. Despite the fact that different libraries display a good correlation between sequencing platforms, qualitative and quantitative variations in the results were found, depending on the protocol used. Thus, when comparing libraries from different biological samples, it is strongly recommended to use the same sequencing platform and protocol in order to ensure the biological relevance of the comparisons. |
format |
article |
author |
Joern Toedling Nicolas Servant Constance Ciaudo Laurent Farinelli Olivier Voinnet Edith Heard Emmanuel Barillot |
author_facet |
Joern Toedling Nicolas Servant Constance Ciaudo Laurent Farinelli Olivier Voinnet Edith Heard Emmanuel Barillot |
author_sort |
Joern Toedling |
title |
Deep-sequencing protocols influence the results obtained in small-RNA sequencing. |
title_short |
Deep-sequencing protocols influence the results obtained in small-RNA sequencing. |
title_full |
Deep-sequencing protocols influence the results obtained in small-RNA sequencing. |
title_fullStr |
Deep-sequencing protocols influence the results obtained in small-RNA sequencing. |
title_full_unstemmed |
Deep-sequencing protocols influence the results obtained in small-RNA sequencing. |
title_sort |
deep-sequencing protocols influence the results obtained in small-rna sequencing. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2012 |
url |
https://doaj.org/article/1b959c8571564678af790e407e572181 |
work_keys_str_mv |
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