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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...

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Autores principales: Rodriguez, Princess D., Mariani, Michael, Gay, Jamie, Hogan, Tyler C., Amiel, Eyal, Deming, Paula B., Frietze, Seth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2021
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.
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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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