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。