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用于脉冲场凝胶电泳的技术和高分辨率DNA大小标记物

Techniques and high resolution DNA size markers for pulsed field gel electrophoresis.

作者信息

Cooney C A

机构信息

Biology Department, Beckman Research Institute, City of Hope, Duarte, CA 91010.

出版信息

Mol Biotechnol. 1994 Oct;2(2):119-27. doi: 10.1007/BF02824804.

DOI:10.1007/BF02824804
PMID:7866871
Abstract

High resolution DNA size markers are described for pulsed field gel electrophoresis (PFGE). These markers provide resolution of 10-20 kbp over a size range from 10 kbp to more than 400 kbp and are produced by partial restriction digestion of lambda phage DNA concatemers (lambda ladder). High resolution markers extending to over 400 kbp are made by partial restriction digestion of lambda ladder embedded in agarose. Detailed methods are described for marker production and for DNA separation by contour-clamped homogeneous electric field (CHEF) electrophoresis. These markers and methods are useful for a variety of high resolution DNA mapping by PFGE.

摘要

本文描述了用于脉冲场凝胶电泳(PFGE)的高分辨率DNA大小标记物。这些标记物在10 kbp至超过400 kbp的大小范围内提供10 - 20 kbp的分辨率,由λ噬菌体DNA串联体(λ阶梯)的部分限制性消化产生。通过对嵌入琼脂糖中的λ阶梯进行部分限制性消化,可制备延伸至超过400 kbp的高分辨率标记物。本文详细描述了标记物制备方法以及通过轮廓夹钳均匀电场(CHEF)电泳进行DNA分离的方法。这些标记物和方法可用于通过PFGE进行的各种高分辨率DNA图谱分析。

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2
Resolution of DNA molecules greater than 5 megabases by contour-clamped homogeneous electric fields.通过轮廓钳位均匀电场解析大于5兆碱基的DNA分子。
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本文引用的文献

1
Preparation, Manipulation, and Pulse Strategy for One-Dimensional Pulsed-Field Gel Electrophoresis (ODPFGE).一维脉冲场凝胶电泳(ODPFGE)的制备、操作及脉冲策略
Methods Mol Biol. 1992;12:73-103. doi: 10.1385/0-89603-229-9:73.
2
Separation and Size Determination of DNA over a 10-,200 kbp Range.
Methods Mol Biol. 1992;12:31-7. doi: 10.1385/0-89603-229-9:31.
3
Resolving Multimegabase DNA Molecules Using Contour-Clamped Homogeneous Electric Fields (CHEF).使用轮廓夹钳均匀电场(CHEF)解析多兆碱基DNA分子。
J Nucleic Acids. 2012;2012:254630. doi: 10.1155/2012/254630. Epub 2012 Feb 12.
Methods Mol Biol. 1992;12:19-30. doi: 10.1385/0-89603-229-9:19.
4
Separation of large DNA molecules by contour-clamped homogeneous electric fields.通过轮廓钳位均匀电场分离大型DNA分子。
Science. 1986 Dec 19;234(4783):1582-5. doi: 10.1126/science.3538420.
5
A novel instrument for separating large DNA molecules with pulsed homogeneous electric fields.一种用于通过脉冲均匀电场分离大型DNA分子的新型仪器。
Science. 1988 Sep 2;241(4870):1203-5. doi: 10.1126/science.3045968.
6
A regularly spaced DNA size standard with 10 kbp resolution for pulsed field gel electrophoresis.一种用于脉冲场凝胶电泳的具有10 kbp分辨率的等间距DNA大小标准品。
Nucleic Acids Res. 1989 Jul 11;17(13):5412. doi: 10.1093/nar/17.13.5412.
7
Plasmid multimers as high resolution molecular weight standards for pulsed field gel electrophoresis.作为脉冲场凝胶电泳高分辨率分子量标准的质粒多聚体
Nucleic Acids Res. 1989 Jul 11;17(13):5413. doi: 10.1093/nar/17.13.5413.
8
Unexpected loss of genomic DNA from agarose gel plugs.
Biotechniques. 1990 Nov;9(5):542, 544, 546-50.