An improved technique for isolating codominant compound microsatellite markers
An improved technique for isolating codominant compound microsatellite markers
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DOI：
10.1007/s10265-006-0274-2
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发表时间：
2006-07-01
影响因子：
2.8
通讯作者：
Hogetsu, Taizo
中科院分区：
文献类型：
--
作者：
Lian, Chunlan;Wadud, Md. Abdul;Hogetsu, Taizo
本文介绍了一种节省大量时间和成本的共显性多态标记(复合微卫星标记)的开发方法。在该技术中，使用复合SSR引物(AC)(6)(AG)(5)或(TC)(6)(AC)(5)和一个适配引物进行抑制-PCR，从构建的DNA文库中扩增出一端有一个复合SSR序列的片段。根据化合物SSR两侧的序列设计了一条基因座特异性引物。以座位特异性和复合SSR两对引物作为复合SSR标记。由于每个标记的设计只需要一个座位特异的引物，而且只需要一个通用的复合SSR引物作为分析所有复合SSR标记的荧光标记，大大降低了开发共显性标记及其多态性分析的成本。我们已经在三裂石竹中演示了这一技术，并容易地开发出11个具有高度多态的共显性标记。利用这项技术成功地分离出其他几种植物的共显性复合SSR标记目前正在进行中。
An approach for developing codominant polymorphic markers (compound microsatellite (SSR) markers), with substantial time and cost savings, is introduced in this paper. In this technique, fragments flanked by a compound SSR sequence at one end were amplified from the constructed DNA library using compound SSR primer (AC)(6)(AG)(5) or (TC)(6)(AC)(5) and an adaptor primer for the suppression-PCR. A locus-specific primer was designed from the sequence flanking the compound SSR. The primer pairs of the locus-specific and compound SSR primers were used as a compound SSR marker. Because only one locus-specific primer was needed for design of each marker and only a common compound SSR primer was needed as the fluorescence-labeled primer for analyzing all the compound SSR markers, this approach substantially reduced the cost of developing codominant markers and analyzing their polymorphism. We have demonstrated this technique for Dendropanax trifidus and easily developed 11 codominant markers with high polymorphism for D. trifidus. Use of the technique for successful isolation of codominant compound SSR markers for several other plant species is currently in progress.