Applicability of an in-house Chelex-100 DNA isolation method for extracting genetic material from sterlet (Acipenser ruthenus) embryos

Abstract

Efficient, cost-effective DNA extraction methods are crucial for molecular research on sturgeon embryos given that substantial sample sizes must frequently be analyzed in short periods of time. The high lipid and carbohydrate contents of sturgeon embryo yolk sacs mean that obtaining genetic material of sufficient quality is challenging. The predominant methods used include tissue/cell lysis, organic extraction, purification on spin columns, and ethanol precipitation. However, these methods are expensive and time-consuming, which significantly limits the throughput of PCR-based molecular analyses. In the present study, we evaluated the usefulness of an in-house Chelex-100 DNA extraction method on sterlet (Acipenser ruthenus) embryos at the neurula developmental stage (48 hours post fertilization) and compared it with two other commercial silica membrane-based kits for isolating genetic material–NucleoSpin Tissue® (Macherey Nagel, Duren, Germany) and Sherlock AX (A&A Biotechnology, Gdynia, Poland). The yield and quality of nucleic acid, its suitability for PCR amplification, and the total cost and complexity of the extraction methods were evaluated and compared. Our results indicated that the in-house Chelex-100 is inexpensive and can be used as an effective high-throughput method of DNA isolation for sterlet embryos.

Keywords

• comparative dna extraction
• dna yield and purity
• cost- and time-efficiency
• pcr amplification quality
• sturgeon genetics