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用于聚丙烯酰胺凝胶电泳分析的轮状病毒双链核糖核酸的快速、简便制备方法。

Rapid, simple method of preparing rotaviral double-stranded ribonucleic acid for analysis by polyacrylamide gel electrophoresis.

作者信息

Theil K W, McCloskey C M, Saif L J, Redman D R, Bohl E H, Hancock D D, Kohler E M, Moorhead P D

出版信息

J Clin Microbiol. 1981 Sep;14(3):273-80. doi: 10.1128/jcm.14.3.273-280.1981.

DOI:10.1128/jcm.14.3.273-280.1981
PMID:6270190
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC271954/
Abstract

A procedure for extracting rotaviral double-stranded ribonucleic acid (RNA) directly from fecal and intestinal specimens collected from calves and pigs is described. This procedure provides a rapid, simple, reproducible method of obtaining rotaviral double-stranded RNA preparations suitable for electrophoretic analysis in polyacrylamide-agarose composite gels. The rotaviral genome electrophoretic migration pattern produced by double-stranded RNA extracted directly from a specimen by this procedure was qualitatively identical to the electrophoretic migration pattern obtained with double-stranded RNA extracted from purified rotavirus derived from the same specimen. Direct extraction of specimens containing porcine rotavirus-like virus by this procedure gave preparations that had electrophoretic migration patterns similar, but not identical, to the characteristic electrophoretic migration pattern of the rotaviral genome. Sufficient rotaviral double-stranded RNA could be extracted from 6 ml of fecal or intestinal specimen by this procedure to permit 15 or more electrophoretic assays.

摘要

本文描述了一种直接从小牛和猪的粪便及肠道标本中提取轮状病毒双链核糖核酸（RNA）的方法。该方法提供了一种快速、简单且可重复的方式，用于获得适用于在聚丙烯酰胺 - 琼脂糖复合凝胶中进行电泳分析的轮状病毒双链RNA制剂。通过此方法直接从标本中提取的双链RNA所产生的轮状病毒基因组电泳迁移模式，在质量上与从同一标本衍生的纯化轮状病毒中提取的双链RNA所获得的电泳迁移模式相同。通过此方法直接提取含有猪轮状病毒样病毒的标本，得到的制剂其电泳迁移模式与轮状病毒基因组的特征电泳迁移模式相似但不完全相同。通过此方法，从6毫升粪便或肠道标本中可提取足够的轮状病毒双链RNA，以进行15次或更多次电泳检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/5ac4366d6e48/jcm00158-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/d52251353b14/jcm00158-0068-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/15a14a646971/jcm00158-0068-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/53f264fec312/jcm00158-0069-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/1bdd7af0e58f/jcm00158-0069-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/5ac4366d6e48/jcm00158-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/d52251353b14/jcm00158-0068-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/15a14a646971/jcm00158-0068-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/53f264fec312/jcm00158-0069-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/1bdd7af0e58f/jcm00158-0069-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bbe/271954/5ac4366d6e48/jcm00158-0070-a.jpg

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A convenient apparatus for vertical gel electrophoresis.一种用于垂直凝胶电泳的便捷装置。
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