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从小鼠精子中分离并鉴定种系DNA。

Isolation and characterization of germ line DNA from mouse sperm.

作者信息

Shiurba R, Nandi S

出版信息

Proc Natl Acad Sci U S A. 1979 Aug;76(8):3947-51. doi: 10.1073/pnas.76.8.3947.

DOI:10.1073/pnas.76.8.3947
PMID:291053
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC383953/
Abstract

Mouse germ line DNA was isolated from sperm by a physicochemical procedure that preferentially destroys contaminating somatic cell DNA. The use of reducing conditions and chelating agents in combination with phenol permitted extraction of molecular weight DNA from mature sperm nuclei with approximately 80% efficiency. Less than 0.1% somatic cell DNA contamination remained in sperm DNA prepared by this method. Germ line DNA was characterized by determination of its ultraviolet absorbance spectrum, buoyant density in cesium chloride, and melting profile on a hydroxyapatite column. Contamination by mitochondrial DNA was assessed by cesium chloride/ethidium bromide gradient centrifugation. The significance of the mouse germ line DNA isolation procedure is discussed with respect to the possible genetic transmission of mammary tumor virus and leukemia virus, the origin of antibody diversity, and the origin of testicular teratomas.

摘要

通过一种优先破坏污染性体细胞DNA的物理化学方法，从精子中分离出小鼠生殖系DNA。在苯酚中使用还原条件和螯合剂，可从成熟精子核中提取分子量DNA，效率约为80%。用这种方法制备的精子DNA中，体细胞DNA污染残留不到0.1%。通过测定其紫外吸收光谱、氯化铯中的浮力密度以及在羟基磷灰石柱上的解链曲线来表征生殖系DNA。通过氯化铯/溴化乙锭梯度离心评估线粒体DNA的污染情况。讨论了小鼠生殖系DNA分离程序对于乳腺肿瘤病毒和白血病病毒可能的遗传传递、抗体多样性的起源以及睾丸畸胎瘤的起源的意义。

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本文引用的文献

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The nucleic acids of plant tissues; the extraction and estimation of desoxypentose nucleic acid and pentose nucleic acid.植物组织的核酸；脱氧戊糖核酸和戊糖核酸的提取与测定
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A new method for the isolation of deoxyribonucleic acids; evidence on the nature of bonds between deoxyribonucleic acid and protein.一种分离脱氧核糖核酸的新方法；关于脱氧核糖核酸与蛋白质之间键的性质的证据。
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A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.一种用于检测和分离闭环双链DNA的染料浮力密度法：HeLa细胞中的闭环DNA
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