Al-Shuhaib Mohammed Baqur S
Department of Animal Production, College of Agriculture, Al-Qasim Green University, 8-Al Qasim, 51001 Babil, Iraq.
J Genet. 2018 Aug 24;97(4):e87-e94.
Here, we describe a rapid, efficient and noncomplicated method for extracting poultry genomic DNA. This simple method obtains excellent qualities of molecular biology grade DNA in a matter of only about 15 min. A straightforward protocol is followed in this extraction procedure, in which, when the blood cells are placed in a distilled water alone, water rapidly disrupts red blood cells (RBCs) membranes and alleviate chickens' undesired high blood viscosity without being aided by any other chemicals. Moreover, the time, cost and efforts were hugely reduced since the step of leukocytes lysis was successfully mixed with the protein precipitation. The isolated genomic DNA, in terms of its quantity and quality, is satisfying and is proved to be suitable for restriction endonucleases digestion, polymerase chain reaction (PCR), and restriction fragment length polymorphism (RFLP). Thus, instead of extracting a limited amount of DNA using expensive or relatively expensive kits, this guaranteed procedure can be utilized alternatively to accurately extract genomic DNA (gDNA) from minimal starter quantities of the chicken's blood that do not exceed 50 L. By relying on this technique, all the practical handover steps are reduced to the extreme limits. This pilot study may provide a high yield gDNA extraction that minimizes labour, expense, and steps in very high competency and reproducibility.
在此,我们描述了一种快速、高效且不复杂的提取家禽基因组DNA的方法。这种简单的方法仅需约15分钟就能获得分子生物学级别的优质DNA。该提取过程遵循一个直接的方案,即当血细胞单独置于蒸馏水中时,水会迅速破坏红细胞(RBC)膜并缓解鸡类不期望的高血液粘度,无需任何其他化学物质的辅助。此外,由于白细胞裂解步骤成功地与蛋白质沉淀步骤合并,时间、成本和工作量大幅减少。就其数量和质量而言,分离出的基因组DNA令人满意,并且被证明适用于限制性内切酶消化、聚合酶链反应(PCR)和限制性片段长度多态性(RFLP)分析。因此,无需使用昂贵或相对昂贵的试剂盒来提取有限量的DNA,这种可靠的方法可用于从不超过50微升的少量鸡血液样本中准确提取基因组DNA(gDNA)。依靠这项技术,所有实际的交接步骤都被减少到了极限。这项初步研究可能提供一种高产的gDNA提取方法,在极高的效率和可重复性下将劳动力、费用和步骤降至最低。