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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| Formato: | article |
| Lenguaje: | EN |
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American Society for Microbiology
2014
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| Acceso en línea: | https://doaj.org/article/caaf4c4916614b94ba8d96d2e8227192 |
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| Sumario: | 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. |
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