蟠桃果实总RNA提取方法的研究

RNA Extraction Method from Mature Flat Peach Fruits

【目的】筛选获得提取蟠桃果肉中总RNA的适宜方法。【方法】以盛花后100 d的蟠桃为材料,采用Transplant plus植物总RNA提取试剂提取法、RNAisoTM试剂盒法、改良SDS法、改良CTAB法等四种方法提取总RNA。【结果】两种试剂盒法提取的总RNA质量差,有严重的基因组污染;改良SDS法提取物中含有大量的多糖;采用改良CTAB法所得RNA完整性好,条带清晰无降解,无基因组污染。经过RT～PCR和Northern blot检测后,表明该RNA能够满足后续分子生物学操作的要求。【结论】蟠桃果实总RNA适宜提取方法-改良CTAR法的确定为后续果实发育分子生物学的研究奠定了基础。

[ Objective ] The target of current research was to select and obtain a proper RNA extraction method from flat peach fruit. [ Method ] RNAplant plus, RNAisoTM, improved SDS method and improved CTAB method were used to isolate total RNA from flat peach fruits harvested at 100 days after full blossom. [ Result ] RNA extracted by two extraction kits had poor quality with contamination of the genome DNA ; RNA extracted by improved SDS method contained large amounts of polysaccharide, while RNA extracted by the improved CTAB method had good integrity and the RNA bands were clear without degradation. RT - PCR and Northern blot test indicated that the extracted RNA could be used for the following molecular biology procedures. [ Conclusion ] The establishment of optimum RNA extraction method from flat peach fruits - improved CTAB method the base for the follow - up molecular biology study of fruit development.