An improved method for the isolation of highly polymerized native deoxyribonucleic acid from certain protozoa.
An improved method for the isolation of highly polymerized native deoxyribonucleic acid from certain protozoa.
复制标题
一种从某些原生动物中分离高度聚合的天然脱氧核糖核酸的改进方法。
DOI：
10.1111/j.1550-7408.1973.tb06021.x
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发表时间：
1973
期刊：
影响因子：
--
通讯作者：
F. Mohn
中科院分区：
文献类型：
--
作者：
B. Honigberg;F. Mohn
概要
一个相对简单的苯酚提取方法，EDTA作为核酸酶抑制剂，描述了用于分离纯化的，高度聚合的天然DNA从阴道毛滴虫，毛滴虫gallinae，和胎儿三毛滴虫，它也适用于梨形四膜虫。RNA酶T1、RNA酶A（Worthington’s R）、链霉蛋白酶和α-淀粉酶digestrin构成获得令人满意的DNA产量的重要步骤。高聚合度的分离产物估计在O.D.260 DNA酶处理后的增色性和氯化铯梯度分析。DNA样品的双链状态通过后一种方法和通过用NaOH变性来估计，并且分子量通过蔗糖梯度分析来估计。样品的纯度通过蛋白质和糖原的化学分析测定。通过二苯胺反应估计DNA回收率。
SYNOPSIS
A relatively simple phenol extraction method, with EDTA as the nuclease inhibitor, is described for the isolation of purified, highly polymerized native DNA from Trichomonas vaginalis, Trichomonas gallinae, and Tritrichomonas foetus; it is applicable also to Tetrahymena pyriformis. RNase Tl, RNase A (Worthington's R), pronase, and α-amylase digestions constitute important steps in obtaining satisfactory yields of DNA. High degree of polymerization of the isolation product was estimated by hyperchromicity at O.D.260 after DNase treatment and by CsCl gradient analysis. The double-stranded condition of the DNA samples was estimated by the latter method and by denaturation with NaOH, and the molecular weight by sucrose gradient analysis. Purity of the samples was determined spectrophotometrically and by chemical analyses for protein and glycogen. DNA percent recovery was estimated by the diphenylamine reaction.