中国水稻科学

• 实验技术 • 上一篇    下一篇

琼脂糖电泳展示mRNA差异方法的探讨

关和新;凌杏元;朱英国   

  1. 武汉大学遗传研究所,武汉 430072
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1998-07-10 发布日期:1998-07-10

Methods of Agarose Gel Electrophoresis for mRNA Differential Display

Guan Hexin; Ling Xingyuan; Zhu Yingguo   

  • Received:1900-01-01 Revised:1900-01-01 Online:1998-07-10 Published:1998-07-10

摘要: 以粤泰不育系和保持系黄化苗及花药为材料,探讨了琼脂糖电泳展示mRNA差异的方法。

关键词: 雄性不育系, 聚合酶链式反应, 恢复系, RNA

Abstract: RT-PCR reactions, with template cDNA from etiolated seedlings of Yuetai maintainer lines (Oryza sativa L. ) and 2, 3 or 4 random primers, were carried out at anneal temperatures 36℃, 38℃, 40℃, 42℃ and45℃. Products were separated on 2% agarose gel. No products were obtained at both 42℃and 45℃. Most of the main bands obtained at 36℃, 38℃ and 40℃ were similar, but there are some differences in the weak bands. Thus, we select 40℃ as the anneal temperature in the following experiments. The difference of mRNAs, which were isolated from anthers at different developmental stages of Yuetai CMS, and maintainer lines, were displayed by agarose gel. Differential bands were retrieved and re-amplified. Then, products were purified on 3% agarose gel. Every purified band showed single band on 5% polyacrylamide gel.

Key words: cytoplasm sterile line, electrophoresis, polymerase chain reaction, restorer line, RNA