Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells

Q3 Agricultural and Biological Sciences

Acta Scientiarum. Biological Sciences Pub Date : 2022-07-27 DOI:10.4025/actascibiolsci.v44i1.59272

A. Santos, Wilson Romão Toledo da Silva, H. N. Ferreira, Antonio Matos Fraga Junior, M. Batista, J. Velarde

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引用次数: 0

Abstract

DNA extraction is usually the first step to perform molecular studies. This process can be nonviable due to genomic DNA (gDNA) extraction commercial kits prices. Furthermore, available DNA extraction protocols generally have high specificity, limiting their use to specific sources of biological material. In order to reduce costs, optimize time and laboratory logistics, besides to demonstrate a versatile protocol, the present study worked on an efficient DNA extraction protocol from somatic and non-somatic cells, using biological material from sheep as a model. For that, gDNA was extracted from whole blood, spermatozoa, and hair bulb cells, collected from three adult sheep, transported at 5ºC and stored at -20ºC until lab procedures. After extraction, gDNA concentration and purity were evaluated in a nano spectrophotometer. gDNA concentration from whole blood was greater (p < 0.05) than extracted from hair bulb cells, which in turn was superior (p < 0.05) than in spermatozoa. Also, gDNA from whole blood and, followed by, sperm showed greater (p < 0.05) purity when compared to gDNA of hair bulb cells. Adapting a gDNA extraction protocol, originally developed for bovine whole blood, enabled to obtain and isolate gDNA in different nucleated sheep cells

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高效、快速、低成本提取不同有核绵羊细胞DNA的方法

DNA提取通常是进行分子研究的第一步。由于基因组DNA (gDNA)提取商业试剂盒的价格，该过程可能不可行。此外，现有的DNA提取方案通常具有高特异性，限制了它们用于特定来源的生物材料。为了降低成本，优化时间和实验室物流，除了展示一种通用的方案外，本研究还以羊的生物材料为模型，研究了一种高效的体细胞和非体细胞DNA提取方案。为此，从3只成年羊的全血、精子和毛球细胞中提取gDNA，在5ºC下运输，在-20ºC下保存，直到进行实验室操作。提取后用纳米分光光度计测定gDNA的浓度和纯度。全血gDNA浓度高于毛球细胞(p < 0.05)，而毛球细胞的gDNA浓度又高于精子(p < 0.05)。与毛球细胞的gDNA相比，全血和精子的gDNA纯度更高(p < 0.05)。采用最初为牛全血开发的gDNA提取方案，能够在不同的有核羊细胞中获得和分离gDNA

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来源期刊

Acta Scientiarum. Biological Sciences Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)

CiteScore

1.00

自引率

0.00%

发文量

审稿时长

38 weeks

期刊介绍： The journal publishes original articles in all areas of Biological Sciences, including anatomy, bacteriology, molecular biology, biochemistry, botany, cytology and cell biology, animal behavior, ecology, limnology, embryology, and histology, morpho-physiology, genetics, microbiology, parasitology and zoology.