摘要
甘蔗是重要的糖料及能源作物之一。甘蔗种质RNA的提取是甘蔗分子生物学研究的前提。以成熟甘蔗的叶片为材料,通过对前人的植物RNA提取方法进行改良,寻求最佳的甘蔗总RNA提取方法。通过改良最后得到的异硫氰酸胍法Ⅲ具有成本低、操作简单、省时等特点。用该法提取得到的甘蔗总RNA经琼脂糖凝胶电泳可获得28S、18S和5SrRNA3条带,并且28SrRNA的亮度大于18SrRNA的亮度。表明该方法提取的RNA完整性好,纯度高。另外对所得部分甘蔗种质的总RNA进行RT-PCR检测,可特异扩增出目标片段,说明用异硫氰酸胍法Ⅲ提取的甘蔗种质总RNA,可以直接用于后续的分子生物学试验。
Sugarcane was one of the most important sugar crops and energy crops in the world. The isolation of Sugarcane RNA was a precondition of Molecular biology research. This experiment took the sugarcane leaves as the material, seeking the best total RNA of sugarcane extraction method through improving the predecessors＇ total RNA extraction methods for plants. The final improved isothiocyanate method Ⅲ had the characteristics of low cost, simple operation, coupled with time-saving and so on. Sugarcane total RNA obtained by this extraction method got the 28S, 18S and 5S rRNA three bands after the agarose gel electrophoresis, and the band of 28S rRNA brighter than 18S RNA. The result indicated that the RNA samples obtained by the method had a good integrity and high purity. In addition, the partial sugarcane total RNA obtained by this method could be increased specially by RT-PCR technique, and obtained the target fragment. The result showed that the RNA which had isolated by isothiocyanate method Ⅲ had highly quality and could be used for the later molecular researches.
出处
《中国农学通报》
CSCD
北大核心
2009年第24期68-72,共5页
Chinese Agricultural Science Bulletin
基金
海南大学科技基金(Rnd0704)
海南省重点学科建设项目(xkxm0811-06)资助
关键词
甘蔗
RNA提取
异硫氰酸胍法
sugarcane, RNA extraction, isothiocyanate method
