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复合聚丙烯酰胺凝胶电泳:一种增加扩增短串联重复序列等位基因分离度的替代方法。

Composite PAGE: an alternate method for increased separation of amplified short tandem repeat alleles.

作者信息

Lorente M, Lorente J A, Wilson M R, Budowle B, Villanueva E

机构信息

Department of Legal Medicine, University of Granada, Spain.

出版信息

Int J Legal Med. 1993;106(2):69-73. doi: 10.1007/BF01225043.

Abstract

Genetic typing of short tandem repeat (STR) loci may require electrophoretic separation techniques which, depending on the locus, can resolve alleles differing in size by only 2, 3 or 4 base pairs (bps). Many such loci can be separated by horizontal, discontinuous polyacrylamide gel electrophoresis (hd-PAGE). However, some loci present particular separation challenges. A composite (or step) gel system consisting of 2 zones, anodal and cathodal, which differ by pore size and ionic strength, has been developed to augment resolution of DNA fragments without a need for additional electrophoretic equipment. Depending on pore size, as well as ionic strength, initial migration can be increased or decreased to effect resolution. When compared with hd-PAGE, this composite gel electrophoretic system can result in enhanced resolution of PCR amplified STR alleles.

摘要

短串联重复序列(STR)基因座的基因分型可能需要电泳分离技术,根据基因座的不同,该技术可以分辨大小仅相差2、3或4个碱基对(bps)的等位基因。许多此类基因座可通过水平不连续聚丙烯酰胺凝胶电泳(hd-PAGE)进行分离。然而,一些基因座存在特殊的分离挑战。一种由阳极和阴极两个区域组成的复合(或阶梯)凝胶系统已被开发出来,这两个区域的孔径和离子强度不同,无需额外的电泳设备即可提高DNA片段的分辨率。根据孔径以及离子强度,可以增加或减少初始迁移以实现分辨率。与hd-PAGE相比,这种复合凝胶电泳系统可以提高PCR扩增的STR等位基因的分辨率。

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