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湖泊科学  2008 

原生动物单细胞PCR应用初探——以天蓝喇叭虫为例

DOI: 10.18307/2008.0517

Keywords: 单细胞PCR,单细胞全基因组扩增,原生动物,天蓝喇叭虫

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Abstract:

以天蓝喇叭虫(Stentorcoeruleus)为研究对象,探索了单细胞单基因PCR扩增及单细胞全基因组PCR扩增技术在原生动物中的应用.经过不断探索和优化条件后,试验取得了理想的结果.在40例单细胞单基因(SSUrDNA基因全序列)PCR的一次性扩增中,新鲜细胞和经过中性红染色的细胞都获得了100%的成功率,室温下酒精(95%)保存一周的细胞获得了82.5%的成功率.在单细胞全基因组PCR扩增中,采用高效高保真的phi29DNA聚合酶结合随机引物(RandomPrimer)进行扩增,获得了丰富且质量较高的PCR产物.以全基因组扩增产物(稀释10倍)对四个常用的基因位点(TEF1、SSUrRNA、18S-ITS1-5.8S、a-tubulin)进行扩增,均成功获得相应的基因片断.

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