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目的:建立金荞麦愈伤组织总RNA提取方法。方法:采用CTAB法结合LiCl沉淀的改良方法提取RNA,紫外分光光度检测、0.8%非变性琼脂糖凝胶电泳检测、RNA的反转录等评价RNA质量。结果:提取的总RNA 28S和18S条带清晰,且A260/A280在1.9～2.0,以提取的RNA为模板反转录的cDNA长度范围较广(500 bp～5 kb)。结论:该方法提取出的总RNA质量能满足RT-PCR等分子生物学操作的要求,且简单、有效、经济,可用于金荞麦愈伤组织总RNA的提取。
Objective: To establish a method for extracting total RNA from callus of callus. Methods: RNA was extracted by CTAB method combined with LiCl precipitation. The RNA was detected by ultraviolet spectrophotometry, 0.8% agarose gel electrophoresis and RNA reverse transcription. Results: The bands of 28S and 18S were clear and the A260 / A280 ranged from 1.9 to 2.0. The length of the reverse transcribed cDNA was about 500 bp ~ 5 kb using the extracted RNA as a template. CONCLUSION: The total RNA extracted by this method can meet the requirements of molecular biology such as RT-PCR and is simple, effective and economical for the extraction of total RNA from callus of Fagopyrums.