淹涝处理下樱桃总RNA提取方法的建立与优化.pdfVIP

上海农业学报 2018,34(2):28-32 ActaAgriculturae Shanghai DOI:10.15955/j.issn1000-3924.2018.02.06 文章编号:1000-3924(2018)02-028-05 淹涝处理下樱桃总RNA提取方法的建立与优化 生利霞,孟祥毅,王 猛,王 倩,冯立国∗ (扬州大学园艺与植物保护学院,扬州225009) 摘 要:以淹涝处理后的樱桃叶片和根系为材料,采用CTAB法、TaKaRa RNA提取试剂盒法和改良试剂盒 法提取其总RNA,通过琼脂糖凝胶电泳和核酸蛋白检测仪比较所提取总 RNA 样品的质量。 结果表明:改良 TaKaRa RNA提取试剂盒法提取的总RNA完整性高,质量好,无降解;进一步采用改良TaKaRa RNA提取试剂盒 法提取樱桃淹涝胁迫0—4 d 叶片和根系的总RNA为模板进行RT-PCR验证,发现PCR扩增得到的条带较清晰, 与目的片段大小一致。 因此,改良TaKaRaRNA提取试剂盒法提取的樱桃叶片与根系总RNA完全可以满足进一 步分子生物学研究的需要,是淹涝处理后樱桃总RNA提取的理想方法。 关键词:樱桃;淹涝处理;总RNA;提取方法 中图分类号:S662.5 文献标识码:A Methods of extracting total RNA from cherry under waterlogging SHENG Li-xia,MENG Xiang-yi,WANG Meng,WANG Qian,FENG Li-guo∗ (School of Horticulture and Plant Protection,Yangzhou University,Yangzhou225009,China) Abstract:Total RNA was isolated from leaves and roots of cherry under waterlogging through CTAB method,TaKaRa MiniBEST Plant RNA Extraction Kit and improved Extraction Kit method.The quality of the total RNA was testedby gel electrophoresisand Ultramicro nucleic acid/protein detector.Theresultsshowedthat total RNA extracted by improvedTaKaRa MiniBEST Plant RNAExtraction Kithadtwo clearbandsof28SrRNA and 18S rRNA without degradation,and with higher purity and good integrity.Then,the total RNA of leaves and roots of cherry after0—4 dayswaterlogging treatment were extracted by improved kit method,the quality of the RNA were determined by RT-PCR,the results showed that the PCR product had a distinct band with the same size as theoretical fragment and could satisfy the demands of molecular biology experiment. So the improved TaKaRa MiniBEST Plant RNA Extraction Kit method was the ideal method for the extraction of total RNA from cherry after waterlogging. Key words:Cherry;Waterlogging treatment;Total RNA;Extraction method 樱桃