T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells

BACKGROUND: RNA sequencing has become the mainstay for studies of gene expression. Still, analysis of rare cells with random hexamer priming – to allow analysis of a broader range of transcripts – remains challenging. RESULTS: We here describe a tagmentation-based, rRNA blocked, random hexamer prime...

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Autores principales: Gustafsson, Charlotte, Hauenstein, Julia, Frengen, Nicolai, Krstic, Aleksandra, Luc, Sidinh, Månsson, Robert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Research
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10111750/
https://www.ncbi.nlm.nih.gov/pubmed/37069502
http://dx.doi.org/10.1186/s12864-023-09279-4
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author Gustafsson, Charlotte
Hauenstein, Julia
Frengen, Nicolai
Krstic, Aleksandra
Luc, Sidinh
Månsson, Robert
author_facet Gustafsson, Charlotte
Hauenstein, Julia
Frengen, Nicolai
Krstic, Aleksandra
Luc, Sidinh
Månsson, Robert
author_sort Gustafsson, Charlotte
collection PubMed
description BACKGROUND: RNA sequencing has become the mainstay for studies of gene expression. Still, analysis of rare cells with random hexamer priming – to allow analysis of a broader range of transcripts – remains challenging. RESULTS: We here describe a tagmentation-based, rRNA blocked, random hexamer primed RNAseq approach (T-RHEX-RNAseq) for generating stranded RNAseq libraries from very low numbers of FACS sorted cells without RNA purification steps. CONCLUSION: T-RHEX-RNAseq provides an easy-to-use, time efficient and automation compatible method for generating stranded RNAseq libraries from rare cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-023-09279-4.
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spelling pubmed-101117502023-04-19 T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells Gustafsson, Charlotte Hauenstein, Julia Frengen, Nicolai Krstic, Aleksandra Luc, Sidinh Månsson, Robert BMC Genomics Research BACKGROUND: RNA sequencing has become the mainstay for studies of gene expression. Still, analysis of rare cells with random hexamer priming – to allow analysis of a broader range of transcripts – remains challenging. RESULTS: We here describe a tagmentation-based, rRNA blocked, random hexamer primed RNAseq approach (T-RHEX-RNAseq) for generating stranded RNAseq libraries from very low numbers of FACS sorted cells without RNA purification steps. CONCLUSION: T-RHEX-RNAseq provides an easy-to-use, time efficient and automation compatible method for generating stranded RNAseq libraries from rare cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-023-09279-4. BioMed Central 2023-04-17 /pmc/articles/PMC10111750/ /pubmed/37069502 http://dx.doi.org/10.1186/s12864-023-09279-4 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Gustafsson, Charlotte
Hauenstein, Julia
Frengen, Nicolai
Krstic, Aleksandra
Luc, Sidinh
Månsson, Robert
T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells
title T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells
title_full T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells
title_fullStr T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells
title_full_unstemmed T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells
title_short T-RHEX-RNAseq – a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells
title_sort t-rhex-rnaseq – a tagmentation-based, rrna blocked, random hexamer primed rnaseq method for generating stranded rnaseq libraries directly from very low numbers of lysed cells
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10111750/
https://www.ncbi.nlm.nih.gov/pubmed/37069502
http://dx.doi.org/10.1186/s12864-023-09279-4
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