効率的マイクロサテライト(SSR)マーカー作製のためのプロトコル(<特集>マイクロサテライトマーカーは森林科学に何をもたらしたか?)  [in Japanese] A Protocol for Efficient Development of Microsatellite Markers(<Special Issue>What Have Microsatellite Markers Presented to Forest Science?)  [in Japanese]

マイクロサテライト(SSR)マーカーは多型性が抜きんでて高く共優性でしかも再現性が高いため,生態学,育種学,集団遺伝学等の分野で重用されている。一方,このマーカーの最大の課題は,まず初めに対象種ごとに特異的なSSRマーカーを作製しなくてはならないことである。従来の方法では,特に,ゲノム中のSSR頻度が少ない種でのマーカー作製には,多大な労力,時間,費用がかかり,大きな障害となっている。ごく最近著者らの研究室では,新たにDual-suppression-PCR法およびISSR-dual-suppression-PCR法を考案し,様々な種で比較的容易かつ効率的にSSRマーカーを作製できるようになった。そこで本稿では,SSR多型解析を志す研究者の便宜を図るべく,そのうち,dual-suppression-PCR法の実践的プロトコルを詳述した。

Because microsatellite (SSR) markers are highly polymorphic, codominant and reproducible, they are widely used as effective genetic markers in many fields such as ecology, breeding and population genetics. One of the major drawbacks of SSR polymorphism analysis is derived from the fact that several species-specific SSR markers must usually be developed for the first time when research is commenced on a species which has never been analyzed before. Especially when the species has SSR loci in the genome at a low frequency, the marker development often requires much labor, time and expense, hindering SSR analysis. Only recently, we have newly established techniques for SSR marker development named dual-suppression-PCR and ISSR-dual-suppression-PCR techniques, and developed much easily and efficiently many SSR markers from many kinds of organisms by these techniques. In this brief review, we described the practical protocol of dual-suppression-PCR technique in detail, for convenience of researchers who challenge SSR marker development.