探讨和田黑鸡淋巴细胞最佳分离条件和淋巴细胞总RNA提取方法,为和田黑鸡淋巴细胞的免疫学研究提供最为有效的实验材料.采用离心法分离鸡外周血淋巴细胞并检测淋巴细胞活力.利用Trizol法提取和田黑鸡的外周血淋巴细胞总RNA,通过琼脂糖凝胶电泳对所提取的RNA进行检测,并用核酸测定仪测定总RNA.结果表明;使用1;1比例的无Ca2+、Mg2+-Hank'S稀释液稀释抗凝血后,在室温、2 000 rpm/min、35 min的条件下离心分离所得的淋巴细胞效果最好.琼脂糖凝胶电泳结果显示出28S、18S和5.8S三条带,使用核酸测定仪测定的总RNA的OD260/280值为1.996,浓度为283 μg/ml,表明提取了高质量的总RNA,为开展后续的研究奠定了坚实的基础.%The experiment investigated the optimum separation conditions and the total RNA extraction method of lymphocyte, which provide the most effective experimental materials for the immunological study of lymphocytes in Hetian black chicken.Lymphocyte was separated by centrifugation and detected the cell activity.The total RNA was extracted from the peripheral blood lymphocytes by Trizol method.The results showed that the best effect of centrifugal separation was 1;1 ratio of Ca2+, Mg2+-Hank'S dilution of anticoagulant, room temperature, 2000 rpm/min, 35 min.Agarose gel electrophoresis showed three band of 28S, 18S and 5.8S,the OD260 / 280 value of Total RNA was 1.996 and the concentration was 283 μg/ml, which indicating that high quality total RNA was extracted, laying a solid foundation for the follow-up studies.
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