DNA ISOLATION FROM BIOLOGICAL MATERIAL OF MARALS

M. Lubennikova, K. A. Afanasiev, V. A. Afanasiev
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Abstract

The main task of maral breeding isproduction increase of antlers. Application of molecular genetic methods in maral breeding contributes to increase in reliability and accuracy of assessment of productivity and breeding value of stag deer at an early age. The purpose of the research is to test methods DNA isolation from biological material of marals. The biomaterial was selected fromstag deer of Altai-Sayan breed (blood, antler crumbs, cartilaginous tissue of the auricles). The work on DNA isolation from maral tissue samples was carried out in the bioengineering laboratory at Altai State University (Barnaul). DNA isolation was conducted using the method based on application of Chelex-100TM Resin chelating agent (Bio-Rad, USA), the method based on DNA precipitation Diamond DNA (OOO ABT, Russia) and commercial kit based on magnetic particles AMPure XP (Beckman Coulter, USA). The purification degree of the isolated DNA was assessed by PCR efficiency in reactions of amplification of the genes of cytochrome oxidase 1, cytochrome B of mitochondrial DNA. The highest concentration of maral DNA was established in isolation based on DNA precipitation (Diamond DNA). The concentration of DNA in the solution was 13.20 ng/µl (blood), 11.30 ng/µl (cartilaginous tissue), 5.74 ng/µl (antler crumbs).
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从动物生物材料中分离DNA
公马繁育的主要任务是增加鹿角产量。分子遗传学方法在鹿群育种中的应用,有助于提高早期鹿群生产力和育种价值评估的可靠性和准确性。本研究的目的是测试从动物生物材料中分离DNA的方法。生物材料选用阿尔泰-萨扬品种鹿血、鹿角屑、耳郭软骨组织。从动物组织样本中分离DNA的工作在阿尔泰国立大学(Barnaul)的生物工程实验室进行。采用基于Chelex-100TM树脂螯合剂(Bio-Rad,美国)的方法,基于DNA沉淀法的Diamond DNA (OOO ABT,俄罗斯)和基于磁性颗粒AMPure XP (Beckman Coulter,美国)的商用试剂盒进行DNA分离。通过线粒体DNA细胞色素氧化酶1、细胞色素B基因扩增反应的PCR效率评价分离DNA的纯化程度。根据DNA沉淀法(Diamond DNA)分离出最高浓度的maral DNA。DNA浓度分别为血液13.20 ng/µl、软骨组织11.30 ng/µl、鹿茸屑5.74 ng/µl。
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