Simple and Effective Squash-PCR for Rapid Genotyping of Industrial Microalgae
Author:
Yuan Guoliang1ORCID, Gao Song2ORCID, Czajka Jeffrey J.1, Dai Ziyu1, Pomraning Kyle R.1, Duong Rylan D.1, Hofstad Beth A.1, Deng Shuang1ORCID
Affiliation:
1. Chemical and Biological Processes Development Group, Pacific Northwest National Laboratory, Richland, WA 99352, USA 2. Marine and Coastal Research Laboratory, Pacific Northwest National Laboratory, Sequim, WA 98382, USA
Abstract
Microalgae are recognized for their versatility in providing renewable energy, biopharmaceuticals, and nutraceuticals, attributed to their sustainable, renewable, and cost-effective nature. Genetic engineering has proven highly effective in enhancing microalgae production. PCR-based genotyping is the primary method for screening genetically transformed microalgae cells. Recently, we developed a novel PCR method, namely Squash-PCR, and employed it for the molecular analysis of industrially important fungi and yeasts. In this study, we successfully implemented the Squash-PCR technique in 12 industrially significant algae species. This approach offers a quick and reliable means of obtaining DNA templates directly from squashed algal cells, eliminating the need for time-consuming and labor-intensive cultivation and genomic DNA extraction steps. Our results demonstrate the effectiveness of Squash-PCR in detecting and characterizing target genes of interest in 12 different algae species. Overall, this study establishes the Squash-PCR method as a valuable tool for molecular studies in algae, enabling researchers to rapidly screen and manipulate genetic traits in diverse algal species.
Funder
Battelle for the Department of Energy U.S. Department of Energy (DOE) Agile Biofoundry
Subject
Paleontology,Space and Planetary Science,General Biochemistry, Genetics and Molecular Biology,Ecology, Evolution, Behavior and Systematics
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