核桃子叶RNA提取方法的研究
Extracting methods of total RNA in cotyledon of walnut.
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摘要: 针对核桃子叶富含酚类、色素、多糖、油脂等物质的特点，以常用的Trizol法、异硫氰酸胍法、SDS法和改良的CTAB法4种方法提取核桃子叶总RNA，通过RNA产率、纯度、电泳图谱来确定适于核桃子叶RNA提取的方法。结果表明：Trizol法和SDS法所提取的RNA纯度较低；异硫氰酸胍法所提取的RNA降解严重，RNA以小分子弥散状分布；改良CTAB法所提取的RNA呈现出28S rRNA、18S rRNA 2条清晰的带，且28S条带宽度是18S条带宽度的2倍左右，其A260/A280比值为1.98，具有很高的纯度，得率为（100±5.31） μg/g。RTPCR实验结果进一步表明，改良CTAB法提取的RNA完全能够用于后续的分子生物学研究。Abstract: Total RNA was isolated from walnut cotyledon by the methods of Trizol, Guanidine Thioeyanate, SDS and improved CTAB，according to walnut cotyledon characteristics of rich phenols, pigment, polysacchride and oil. The method of RNA isolation suitable for cotyledon of walnut was established through the rate of yield，purity and electrophoresis. The results showed that the purity of RNA isolated by Trizol and SDS reagent was lower than improved CTAB. RNA isolated by the Guanidine Thioeyanate dispersed in the pattern of small molecular. RNA isolated by the improved CTAB showed two clear bands28S rRNA, 18S rRNA respectively and the 28S was two times length as 18S, which means little RNA is decomposed by the method of improved CTAB reagent and the purity of RNA is very high. Yield rate of RNA extracted by the improved CTAB was (100±5.31） μg/g. Its ratio of A260/A280 was 1.98RTPCR amplification also showed that the quality of RNA isolated from the improved CTAB can meet the demands of molecular biology experiment.
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Keywords:
- walnut /
- cotyledon /
- isolation of RNA /
- improved CTAB methods
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