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A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course
Next Generation Sequencing (NGS) has become an important tool in the biological sciences and has a growing number of applications across medical fields. Currently, few undergraduate programs provide training in the design and implementation of NGS applications. Here, we describe an inquiry‐based lab...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley & Sons, Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8356555/ https://www.ncbi.nlm.nih.gov/pubmed/33939256 http://dx.doi.org/10.1002/bmb.21514 |
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author | Rodriguez, Princess D. Mariani, Michael Gay, Jamie Hogan, Tyler C. Amiel, Eyal Deming, Paula B. Frietze, Seth |
author_facet | Rodriguez, Princess D. Mariani, Michael Gay, Jamie Hogan, Tyler C. Amiel, Eyal Deming, Paula B. Frietze, Seth |
author_sort | Rodriguez, Princess D. |
collection | PubMed |
description | Next Generation Sequencing (NGS) has become an important tool in the biological sciences and has a growing number of applications across medical fields. Currently, few undergraduate programs provide training in the design and implementation of NGS applications. Here, we describe an inquiry‐based laboratory exercise for a college‐level molecular biology laboratory course that uses real‐time MinION deep sequencing and bioinformatics to investigate characteristic genetic variants found in cancer cell‐lines. The overall goal for students was to identify non‐small cell lung cancer (NSCLC) cell‐lines based on their unique genomic profiles. The units described in this laboratory highlight core principles in multiplex PCR primer design, real‐time deep sequencing, and bioinformatics analysis for genetic variants. We found that the MinION device is an appropriate, feasible tool that provides a comprehensive, hands‐on NGS experience for undergraduates. Student evaluations demonstrated increased confidence in using molecular techniques and enhanced understanding of NGS concepts. Overall, this exercise provides a pedagogical tool for incorporating NGS approaches in the teaching laboratory as way of enhancing students' comprehension of genomic sequence analysis. Further, this NGS lab module can easily be added to a variety of lab‐based courses to help undergraduate students learn current DNA sequencing methods with limited effort and cost. |
format | Online Article Text |
id | pubmed-8356555 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley & Sons, Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-83565552021-08-11 A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course Rodriguez, Princess D. Mariani, Michael Gay, Jamie Hogan, Tyler C. Amiel, Eyal Deming, Paula B. Frietze, Seth Biochem Mol Biol Educ Articles Next Generation Sequencing (NGS) has become an important tool in the biological sciences and has a growing number of applications across medical fields. Currently, few undergraduate programs provide training in the design and implementation of NGS applications. Here, we describe an inquiry‐based laboratory exercise for a college‐level molecular biology laboratory course that uses real‐time MinION deep sequencing and bioinformatics to investigate characteristic genetic variants found in cancer cell‐lines. The overall goal for students was to identify non‐small cell lung cancer (NSCLC) cell‐lines based on their unique genomic profiles. The units described in this laboratory highlight core principles in multiplex PCR primer design, real‐time deep sequencing, and bioinformatics analysis for genetic variants. We found that the MinION device is an appropriate, feasible tool that provides a comprehensive, hands‐on NGS experience for undergraduates. Student evaluations demonstrated increased confidence in using molecular techniques and enhanced understanding of NGS concepts. Overall, this exercise provides a pedagogical tool for incorporating NGS approaches in the teaching laboratory as way of enhancing students' comprehension of genomic sequence analysis. Further, this NGS lab module can easily be added to a variety of lab‐based courses to help undergraduate students learn current DNA sequencing methods with limited effort and cost. John Wiley & Sons, Inc. 2021-05-03 2021 /pmc/articles/PMC8356555/ /pubmed/33939256 http://dx.doi.org/10.1002/bmb.21514 Text en © 2021 The Authors. Biochemistry and Molecular Biology Education published by Wiley Periodicals LLC on behalf of International Union of Biochemistry and Molecular Biology https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Articles Rodriguez, Princess D. Mariani, Michael Gay, Jamie Hogan, Tyler C. Amiel, Eyal Deming, Paula B. Frietze, Seth A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
title | A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
title_full | A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
title_fullStr | A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
title_full_unstemmed | A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
title_short | A guided‐inquiry investigation of genetic variants using Oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
title_sort | guided‐inquiry investigation of genetic variants using oxford nanopore sequencing for an undergraduate molecular biology laboratory course |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8356555/ https://www.ncbi.nlm.nih.gov/pubmed/33939256 http://dx.doi.org/10.1002/bmb.21514 |
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