Cargando…

Evaluation of a high-throughput, cost-effective Illumina library preparation kit

Library preparation for high-throughput sequencing applications is a critical step in producing representative, unbiased sequencing data. The iGenomX Riptide High Throughput Rapid Library Prep Kit purports to provide high-quality sequencing data with lower costs compared to other Illumina library ki...

Descripción completa

Detalles Bibliográficos
Autores principales: Tvedte, Eric S., Michalski, Jane, Cheng, Shaoji, Patkus, Rayanna S., Tallon, Luke J., Sadzewicz, Lisa, Bruno, Vincent M., Silva, Joana C., Rasko, David A., Dunning Hotopp, Julie C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8342411/
https://www.ncbi.nlm.nih.gov/pubmed/34354114
http://dx.doi.org/10.1038/s41598-021-94911-0
_version_ 1783734063590277120
author Tvedte, Eric S.
Michalski, Jane
Cheng, Shaoji
Patkus, Rayanna S.
Tallon, Luke J.
Sadzewicz, Lisa
Bruno, Vincent M.
Silva, Joana C.
Rasko, David A.
Dunning Hotopp, Julie C.
author_facet Tvedte, Eric S.
Michalski, Jane
Cheng, Shaoji
Patkus, Rayanna S.
Tallon, Luke J.
Sadzewicz, Lisa
Bruno, Vincent M.
Silva, Joana C.
Rasko, David A.
Dunning Hotopp, Julie C.
author_sort Tvedte, Eric S.
collection PubMed
description Library preparation for high-throughput sequencing applications is a critical step in producing representative, unbiased sequencing data. The iGenomX Riptide High Throughput Rapid Library Prep Kit purports to provide high-quality sequencing data with lower costs compared to other Illumina library kits. To test these claims, we compared sequence data quality of Riptide libraries to libraries constructed with KAPA Hyper and NEBNext Ultra. Across several single-source genome samples, mapping performance and de novo assembly of Riptide libraries were similar to conventional libraries prepared with the same DNA. Poor performance of some libraries resulted in low sequencing depth. In particular, degraded DNA samples may be challenging to sequence with Riptide. There was little cross-well plate contamination with the overwhelming majority of reads belong to the proper source genomes. The sequencing of metagenome samples using different Riptide primer sets resulted in variable taxonomic assignment of reads. Increased adoption of the Riptide kit will decrease library preparation costs. However, this method might not be suitable for degraded DNA.
format Online
Article
Text
id pubmed-8342411
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-83424112021-08-06 Evaluation of a high-throughput, cost-effective Illumina library preparation kit Tvedte, Eric S. Michalski, Jane Cheng, Shaoji Patkus, Rayanna S. Tallon, Luke J. Sadzewicz, Lisa Bruno, Vincent M. Silva, Joana C. Rasko, David A. Dunning Hotopp, Julie C. Sci Rep Article Library preparation for high-throughput sequencing applications is a critical step in producing representative, unbiased sequencing data. The iGenomX Riptide High Throughput Rapid Library Prep Kit purports to provide high-quality sequencing data with lower costs compared to other Illumina library kits. To test these claims, we compared sequence data quality of Riptide libraries to libraries constructed with KAPA Hyper and NEBNext Ultra. Across several single-source genome samples, mapping performance and de novo assembly of Riptide libraries were similar to conventional libraries prepared with the same DNA. Poor performance of some libraries resulted in low sequencing depth. In particular, degraded DNA samples may be challenging to sequence with Riptide. There was little cross-well plate contamination with the overwhelming majority of reads belong to the proper source genomes. The sequencing of metagenome samples using different Riptide primer sets resulted in variable taxonomic assignment of reads. Increased adoption of the Riptide kit will decrease library preparation costs. However, this method might not be suitable for degraded DNA. Nature Publishing Group UK 2021-08-05 /pmc/articles/PMC8342411/ /pubmed/34354114 http://dx.doi.org/10.1038/s41598-021-94911-0 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This 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/) .
spellingShingle Article
Tvedte, Eric S.
Michalski, Jane
Cheng, Shaoji
Patkus, Rayanna S.
Tallon, Luke J.
Sadzewicz, Lisa
Bruno, Vincent M.
Silva, Joana C.
Rasko, David A.
Dunning Hotopp, Julie C.
Evaluation of a high-throughput, cost-effective Illumina library preparation kit
title Evaluation of a high-throughput, cost-effective Illumina library preparation kit
title_full Evaluation of a high-throughput, cost-effective Illumina library preparation kit
title_fullStr Evaluation of a high-throughput, cost-effective Illumina library preparation kit
title_full_unstemmed Evaluation of a high-throughput, cost-effective Illumina library preparation kit
title_short Evaluation of a high-throughput, cost-effective Illumina library preparation kit
title_sort evaluation of a high-throughput, cost-effective illumina library preparation kit
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8342411/
https://www.ncbi.nlm.nih.gov/pubmed/34354114
http://dx.doi.org/10.1038/s41598-021-94911-0
work_keys_str_mv AT tvedteerics evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT michalskijane evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT chengshaoji evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT patkusrayannas evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT tallonlukej evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT sadzewiczlisa evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT brunovincentm evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT silvajoanac evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT raskodavida evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit
AT dunninghotoppjuliec evaluationofahighthroughputcosteffectiveilluminalibrarypreparationkit