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鸡(家鸡)中AFLP标记的多色荧光检测与图谱绘制

Multicolour fluorescent detection and mapping of AFLP markers in chicken (Gallus domesticus).

作者信息

Herbergs J, Siwek M, Crooijmans R P, Van der Poel J J, Groenen M A

机构信息

Animal Breeding and Genetics Group, Wageningen Institute of Animal Sciences, Wageningen Agricultural University, The Netheralnds.

出版信息

Anim Genet. 1999 Aug;30(4):274-85. doi: 10.1046/j.1365-2052.1999.00494.x.

Abstract

We describe the mapping of amplified restriction fragment polymorphism (AFLP) markers in chicken (Gallus domesticus) using a multi-colour fluorescent detection system. DNA was used from a population consisting of four families with a total of 183 F2 individuals. The enzyme combination EcoRI/TaqI was used for double digestion, and fluorescently labelled fragments were analysed on an ABI PRISM 377 DNA sequencer. Polymorphic signals in the range of 50-500 bp were genotyped with the ABI PRISM Genotyper 2.0 software, which enabled the analysis of both dominant and incomplete dominant markers (with respect to AFLP, often referred to as codominant). In 19 sets consisting of 3 EcoRI/TaqI primer pair combinations each, a total of 475 polymorphic markers was detected. From these polymorphisms 344 markers could be mapped on the Wageningen linkage map. Fourteen markers were length polymorphisms of the same fragment and 28 markers Z-linked and uniformative; 64 AFLP markers appeared to be unlinked and 25 AFLP markers could not be accurately mapped on the basis of the genotyping results. The resulting AFLP/microsatellite linkage map is comprised of 33 linkage groups with a total of 835 loci.

摘要

我们描述了使用多色荧光检测系统对鸡(家鸡)的扩增限制性片段多态性(AFLP)标记进行的图谱绘制。DNA取自一个由四个家系组成的群体,共有183个F2个体。酶组合EcoRI/TaqI用于双酶切,荧光标记的片段在ABI PRISM 377 DNA测序仪上进行分析。利用ABI PRISM Genotyper 2.0软件对50 - 500 bp范围内的多态性信号进行基因分型,该软件能够分析显性和不完全显性标记(就AFLP而言,通常称为共显性)。在由19组组成的每组包含3个EcoRI/TaqI引物对组合中,共检测到475个多态性标记。从这些多态性中,344个标记可以定位到瓦赫宁根连锁图谱上。14个标记是同一片段的长度多态性,28个标记是Z连锁且无信息的;64个AFLP标记似乎不连锁,25个AFLP标记根据基因分型结果无法准确定位。所得的AFLP/微卫星连锁图谱由33个连锁群组成,共有835个位点。

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