Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs

Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs

Summary: The structure of 5′ untranslated regions (5′ UTRs) of bacterial mRNAs often determines the fate of the transcripts. Using a dimethyl sulfate mutational profiling with sequencing (DMS-MaPseq) approach, we developed a protocol to generate sequence libraries to determine the base-pairing statu...

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Main Authors: Dmitriy Ignatov, Karolis Vaitkevicius, Jörgen Johansson
Format: Article
Language:English
Published: Elsevier 2020-09-01
Series:STAR Protocols
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166720300332
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spelling doaj-578b032541ed4cb38ac6bd75b155c10d2020-11-25T04:09:05ZengElsevierSTAR Protocols2666-16672020-09-0112100046Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRsDmitriy Ignatov0Karolis Vaitkevicius1Jörgen Johansson2Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research, Umeå University, Umeå, Sweden; Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden; Corresponding authorDepartment of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research, Umeå University, Umeå, Sweden; Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, SwedenDepartment of Molecular Biology, Umeå University, 901 87 Umeå, Sweden; Umeå Centre for Microbial Research, Umeå University, Umeå, Sweden; Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden; Corresponding authorSummary: The structure of 5′ untranslated regions (5′ UTRs) of bacterial mRNAs often determines the fate of the transcripts. Using a dimethyl sulfate mutational profiling with sequencing (DMS-MaPseq) approach, we developed a protocol to generate sequence libraries to determine the base-pairing status of adenines and cytosines in the 5′ UTRs of bacterial mRNAs. Our method increases the sequencing depth of the 5′ UTRs and allows detection of changes in their structures by sequencing libraries of moderate sizes.For complete details on the use and execution of this protocol, please refer to Ignatov et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166720300332
collection DOAJ
language English
format Article
sources DOAJ
author Dmitriy Ignatov
Karolis Vaitkevicius
Jörgen Johansson
spellingShingle Dmitriy Ignatov
Karolis Vaitkevicius
Jörgen Johansson
Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs
STAR Protocols
author_facet Dmitriy Ignatov
Karolis Vaitkevicius
Jörgen Johansson
author_sort Dmitriy Ignatov
title Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs
title_short Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs
title_full Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs
title_fullStr Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs
title_full_unstemmed Generation of Sequencing Libraries for Structural Analysis of Bacterial 5′ UTRs
title_sort generation of sequencing libraries for structural analysis of bacterial 5′ utrs
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2020-09-01
description Summary: The structure of 5′ untranslated regions (5′ UTRs) of bacterial mRNAs often determines the fate of the transcripts. Using a dimethyl sulfate mutational profiling with sequencing (DMS-MaPseq) approach, we developed a protocol to generate sequence libraries to determine the base-pairing status of adenines and cytosines in the 5′ UTRs of bacterial mRNAs. Our method increases the sequencing depth of the 5′ UTRs and allows detection of changes in their structures by sequencing libraries of moderate sizes.For complete details on the use and execution of this protocol, please refer to Ignatov et al. (2020).
url http://www.sciencedirect.com/science/article/pii/S2666166720300332
work_keys_str_mv AT dmitriyignatov generationofsequencinglibrariesforstructuralanalysisofbacterial5utrs
AT karolisvaitkevicius generationofsequencinglibrariesforstructuralanalysisofbacterial5utrs
AT jorgenjohansson generationofsequencinglibrariesforstructuralanalysisofbacterial5utrs
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