应用SMARTercDNA合成技术扩增落地生根叶片的总RNA

摘要

以落地生根(Kalanchoe daigremontiana)未长芽和长芽的叶片为材料,提取落地生根的总RNA,采用SMARTer cDNA合成技术合成cDNA第1链,优化扩增第2链,通过设置梯度PCR扩增循环数,电泳分析后鉴定cDNA质量.结果表明:未长芽和长芽的材料各1μg总RNA分别成功扩增质量较高的双链cDNA.此方法的应用解决了研究材料有限的问题,可由微量的总RNA扩增出足够多的高质量双链cDNA,为进一步从基因水平研究落地生根奠定基础.%Total RNA was extracted from Kalanchoe daigremontiana leaves with or without bulbiferous spurs. The first-strand of cDNA synthesis and the double-strands cDNA amplification were performed u-sing super SMARTer cDNA synthesis technology. The quality of cDNA was evaluated through the gradient of PCR amplification cycles and electrophoresis. High quality double-strands cDNA were obtained from 1 ug total RNA. This method can solve the problem of limited research materials as sufficient high-quality double-strands cDNA are obtained from small amounts of total RNA. This technique allows further research of Kalanchoe daigremontiana.