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Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator
Removal of excess dideoxy terminators from the sequencing mix after the enzymatic reaction is a key process affecting the dideoxy/Sanger sequencing quality. Ethanol precipitation may be the most popular clean-up method because of its low cost; however, it takes a long centrifugation time and frequen...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9778496/ https://www.ncbi.nlm.nih.gov/pubmed/36548368 http://dx.doi.org/10.1371/journal.pone.0279432 |
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author | Sassa, Hidenori Ikebe, Kota |
author_facet | Sassa, Hidenori Ikebe, Kota |
author_sort | Sassa, Hidenori |
collection | PubMed |
description | Removal of excess dideoxy terminators from the sequencing mix after the enzymatic reaction is a key process affecting the dideoxy/Sanger sequencing quality. Ethanol precipitation may be the most popular clean-up method because of its low cost; however, it takes a long centrifugation time and frequently results in low quality sequence data. Commercially available clean-up kits provide high quality sequence data, while they generally have high cost. Here, we describe rapid, effective and low-cost dideoxy terminator clean-up method using a home-made magnetic beads suspension, MagNA, and magnetic separator. We found that MagNA enables rapid and efficient clean-up at ~1/100 of the cost of commercially available kits. The magnetic separator made using low-cost neodymium magnets worked well for the MagNA separation, representing a rapid, efficient and cost-effective dideoxy terminator clean-up system. |
format | Online Article Text |
id | pubmed-9778496 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-97784962022-12-23 Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator Sassa, Hidenori Ikebe, Kota PLoS One Lab Protocol Removal of excess dideoxy terminators from the sequencing mix after the enzymatic reaction is a key process affecting the dideoxy/Sanger sequencing quality. Ethanol precipitation may be the most popular clean-up method because of its low cost; however, it takes a long centrifugation time and frequently results in low quality sequence data. Commercially available clean-up kits provide high quality sequence data, while they generally have high cost. Here, we describe rapid, effective and low-cost dideoxy terminator clean-up method using a home-made magnetic beads suspension, MagNA, and magnetic separator. We found that MagNA enables rapid and efficient clean-up at ~1/100 of the cost of commercially available kits. The magnetic separator made using low-cost neodymium magnets worked well for the MagNA separation, representing a rapid, efficient and cost-effective dideoxy terminator clean-up system. Public Library of Science 2022-12-22 /pmc/articles/PMC9778496/ /pubmed/36548368 http://dx.doi.org/10.1371/journal.pone.0279432 Text en © 2022 Sassa, Ikebe https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Lab Protocol Sassa, Hidenori Ikebe, Kota Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
title | Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
title_full | Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
title_fullStr | Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
title_full_unstemmed | Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
title_short | Rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
title_sort | rapid, effective and low-cost purification of dideoxy-sequencing reactions by home-made magnetic beads suspension and magnetic separator |
topic | Lab Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9778496/ https://www.ncbi.nlm.nih.gov/pubmed/36548368 http://dx.doi.org/10.1371/journal.pone.0279432 |
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