摘要
【目的】建立一种有效的巴旦木花药总RNA提取方法,为巴旦木花粉SFB基因克隆和自交不亲和机制研究奠定基础。【方法】利用改良的CTAB法从巴旦木（Amygdalus communis L.）花药中提取高质量的总RNA。【结果】该方法获得的巴旦木花药总RNA完整性好、无多糖和蛋白质的污染,产量较高（鲜花药为283.44μg/g）,OD260/OD280在1.89-1.99,OD260/OD230〉2.0。提取的巴旦木花药总RNA反转录成cDNA,以SFB（S-haplotype-specific F-box gene）基因简并引物PCR扩增,获得了目的片段。【结论】改良的CTAB法是一种简单、快速有效的巴旦木花药总RNA提取方法,提取的花药总RNA能够满足巴旦木进一步的分子生物学研究。
【Objective】The research aimed to set up an effective method of total RNA extraction from anther of almond（Amygdalus communis L.） and to provide basis for SFB gene（S-haplotype-specific F-box gene）clone and self-incompatibile research.【Method】The improved CTAB method was used to isolate with total RNA from almond anther with.【Result】The result showed that the method could obtain the total RNA with good integrity and without stain of polysaccharide and protein,whose yield was 283.44 μg/g（fresh pollen）.Its OD260/OD280 was between 1.89 and 1.99,OD260/OD230〉2.0.Extracted total RNA from almond anther generated cDNA through reverse transcription reaction,then PCR amplification gained the aimed DNA fragment directed with degenerate primer of SFB gene.【Conclusion】The improved CTAB method is a easy,quick and valid method for total RNA extraction from almond anther.The obtained total RNA is able to to be used for molecular biology study of almond.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2010年第5期898-901,共4页
Xinjiang Agricultural Sciences
基金
国家自然科学基金项目(30760146)
国家科技支撑项目(2007BAD36B01)
