Cargando…

High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer

Recently, Pacific Biosciences released a new highly accurate long-read sequencer called the Revio System that is projected to generate 30× HiFi whole-genome sequencing for the human genome within one sequencing SMRT Cell. Mouse and human genomes are similar in size. In this study, we sought to test...

Descripción completa

Detalles Bibliográficos
Autores principales: Manuel, Juana G., Heins, Hillary B., Crocker, Sandra, Neidich, Julie A., Sadzewicz, Lisa, Tallon, Luke, Turner, Tychele N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10274723/
https://www.ncbi.nlm.nih.gov/pubmed/37333171
http://dx.doi.org/10.1101/2023.06.06.543940
_version_ 1785059786030579712
author Manuel, Juana G.
Heins, Hillary B.
Crocker, Sandra
Neidich, Julie A.
Sadzewicz, Lisa
Tallon, Luke
Turner, Tychele N.
author_facet Manuel, Juana G.
Heins, Hillary B.
Crocker, Sandra
Neidich, Julie A.
Sadzewicz, Lisa
Tallon, Luke
Turner, Tychele N.
author_sort Manuel, Juana G.
collection PubMed
description Recently, Pacific Biosciences released a new highly accurate long-read sequencer called the Revio System that is projected to generate 30× HiFi whole-genome sequencing for the human genome within one sequencing SMRT Cell. Mouse and human genomes are similar in size. In this study, we sought to test this new sequencer by characterizing the genome and epigenome of the mouse neuronal cell line Neuro-2a. We generated long-read HiFi whole-genome sequencing on three Revio SMRT Cells, achieving a total coverage of 98×, with 30×, 32×, and 36× coverage respectively for each of the three Revio SMRT Cells. We performed several tests on these data including single-nucleotide variant and small insertion detection using GPU-accelerated DeepVariant, structural variant detection with pbsv, methylation detection with pb-CpG-tools, and generating de novo assemblies with the HiCanu and hifiasm assemblers. Overall, we find consistency across SMRT Cells in coverage, detection of variation, methylation, and de novo assemblies for each of the three SMRT Cells.
format Online
Article
Text
id pubmed-10274723
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Cold Spring Harbor Laboratory
record_format MEDLINE/PubMed
spelling pubmed-102747232023-06-17 High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer Manuel, Juana G. Heins, Hillary B. Crocker, Sandra Neidich, Julie A. Sadzewicz, Lisa Tallon, Luke Turner, Tychele N. bioRxiv Article Recently, Pacific Biosciences released a new highly accurate long-read sequencer called the Revio System that is projected to generate 30× HiFi whole-genome sequencing for the human genome within one sequencing SMRT Cell. Mouse and human genomes are similar in size. In this study, we sought to test this new sequencer by characterizing the genome and epigenome of the mouse neuronal cell line Neuro-2a. We generated long-read HiFi whole-genome sequencing on three Revio SMRT Cells, achieving a total coverage of 98×, with 30×, 32×, and 36× coverage respectively for each of the three Revio SMRT Cells. We performed several tests on these data including single-nucleotide variant and small insertion detection using GPU-accelerated DeepVariant, structural variant detection with pbsv, methylation detection with pb-CpG-tools, and generating de novo assemblies with the HiCanu and hifiasm assemblers. Overall, we find consistency across SMRT Cells in coverage, detection of variation, methylation, and de novo assemblies for each of the three SMRT Cells. Cold Spring Harbor Laboratory 2023-06-07 /pmc/articles/PMC10274723/ /pubmed/37333171 http://dx.doi.org/10.1101/2023.06.06.543940 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Manuel, Juana G.
Heins, Hillary B.
Crocker, Sandra
Neidich, Julie A.
Sadzewicz, Lisa
Tallon, Luke
Turner, Tychele N.
High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer
title High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer
title_full High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer
title_fullStr High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer
title_full_unstemmed High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer
title_short High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer
title_sort high coverage highly accurate long-read sequencing of a mouse neuronal cell line using the pacbio revio sequencer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10274723/
https://www.ncbi.nlm.nih.gov/pubmed/37333171
http://dx.doi.org/10.1101/2023.06.06.543940
work_keys_str_mv AT manueljuanag highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer
AT heinshillaryb highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer
AT crockersandra highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer
AT neidichjuliea highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer
AT sadzewiczlisa highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer
AT tallonluke highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer
AT turnertychelen highcoveragehighlyaccuratelongreadsequencingofamouseneuronalcelllineusingthepacbioreviosequencer