›› 2011, Vol. 19 ›› Issue (6): 1051-1054.DOI: 10.11733/j.issn.1007-0435.2011.06.027

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Total RNA Amplification from Leaves of Kalanchoe daigremontiana Using SMARTer cDNA Synthesis Technology

ZHONG Tian-xiu, ZENG Hui-ming   

  1. Institute of Turf Grass Science of Beijing Forestry University, Beijing 100083, China
  • Received:2011-04-06 Revised:2011-06-20 Online:2011-12-15 Published:2012-07-09

应用SMARTer cDNA合成技术扩增落地生根叶片的总RNA

钟天秀, 曾会明   

  1. 北京林业大学草坪研究所, 北京100083
  • 通讯作者: 曾会明,E-mail:sciinfo@bjfu.edu.cn
  • 作者简介:钟天秀(1986- ),女,四川内江人,硕士研究生,研究方向为草坪草生物技术育种,E-mail:zhongxinbi@163.com
  • 基金资助:
    “十一五”国家863计划项目(2009AA10Z109)资助

Abstract: 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 using 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.

Key words: RNA, SMARTer cDNA synthesis technology, Kalanchoe daigremontiana

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SMAR-Ter 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。

关键词: RNA, SMARTer cDNA合成, 落地生根

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