Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory

A yeast identification procedure based on the sequencing of the D1/D2 region of the yeast 26S ribosomal DNA was presented in an undergraduate general mycology laboratory. Extracted genomic DNA of environmental yeasts collected by the students was used to PCR-amplify the D1/D2 region of the isolates....

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Autor principal:	William W. Safranek
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Special aspects of education LC8-6691 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/caaf4c4916614b94ba8d96d2e8227192
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spelling	oai:doaj.org-article:caaf4c4916614b94ba8d96d2e82271922021-11-15T15:03:37ZYeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory10.1128/jmbe.v15i1.5981935-78851935-7877https://doaj.org/article/caaf4c4916614b94ba8d96d2e82271922014-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.v15i1.598https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885A yeast identification procedure based on the sequencing of the D1/D2 region of the yeast 26S ribosomal DNA was presented in an undergraduate general mycology laboratory. Extracted genomic DNA of environmental yeasts collected by the students was used to PCR-amplify the D1/D2 region of the isolates. Agarose gel electrophoresis confirmed the presence of the expected amplicon which was purified for sequencing by a spincolumn method then submitted to a commercial sequencing laboratory. The students examined the sequence of their isolates for base miscalls using Applied Biosytem’s Sequence Scan program. The sequences were then used to perform a BLAST search which compared them to the D1/D2 sequences of all validly described yeast species on file in the GenBank database. Of 37 isolates tested, 33 (89%) returned usable sequences for BLAST identification. The module required two lab periods of 1.5 and 2 hours, respectively, to complete with an enrollment of 40 students.William W. SafranekAmerican Society for MicrobiologyarticleSpecial aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 15, Iss 1, Pp 26-27 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Special aspects of education LC8-6691 Biology (General) QH301-705.5
spellingShingle	Special aspects of education LC8-6691 Biology (General) QH301-705.5 William W. Safranek Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory
description	A yeast identification procedure based on the sequencing of the D1/D2 region of the yeast 26S ribosomal DNA was presented in an undergraduate general mycology laboratory. Extracted genomic DNA of environmental yeasts collected by the students was used to PCR-amplify the D1/D2 region of the isolates. Agarose gel electrophoresis confirmed the presence of the expected amplicon which was purified for sequencing by a spincolumn method then submitted to a commercial sequencing laboratory. The students examined the sequence of their isolates for base miscalls using Applied Biosytem’s Sequence Scan program. The sequences were then used to perform a BLAST search which compared them to the D1/D2 sequences of all validly described yeast species on file in the GenBank database. Of 37 isolates tested, 33 (89%) returned usable sequences for BLAST identification. The module required two lab periods of 1.5 and 2 hours, respectively, to complete with an enrollment of 40 students.
format	article
author	William W. Safranek
author_facet	William W. Safranek
author_sort	William W. Safranek
title	Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory
title_short	Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory
title_full	Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory
title_fullStr	Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory
title_full_unstemmed	Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory
title_sort	yeast identification by dna sequencing in an undergraduate mycology laboratory
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/caaf4c4916614b94ba8d96d2e8227192
work_keys_str_mv	AT williamwsafranek yeastidentificationbydnasequencinginanundergraduatemycologylaboratory
_version_	1718428340793114624

Yeast Identification by DNA Sequencing in an Undergraduate Mycology Laboratory

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