檀香心材总RNA提取方法的比较研究  

Comparison of Several RNA Extracting Methods from Heartwood of Santalum album

作　　者：牛美云 程庆伟[1,2] 张月雅[1,2] 张新华[1] 马国华[1] NIU Mei-yun;CHENG Qing-wei;ZHANG Yue-ya;ZHANG Xin-hua;MA Guo-hua(South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China;University of Chinese Academy of Sciences, Beijing 10039,China)

机构地区：[1]中国科学院华南植物园,广州510650 [2]中国科学院大学,北京100049

出　　处：《热带亚热带植物学报》2017年第1期20-25,共6页Journal of Tropical and Subtropical Botany

基　　金：国家自然科学基金项目(31270720;31100498);广东省自然科学基金项目(S2012010009025);广东省科技项目(2015B020231008);中国科学院STS项目子课题(KFJ-EW-STS-118)资助~~

摘　　要：为筛选檀香心材总RNA提取方法,对5种提取方法进行比较研究,包括Trizol法、改良CTAB法、SDS酸酚法、异硫氰酸胍-CTAB法、异硫氰酸胍-SDS法。结果表明,Trizol法和异硫氰酸胍-CTAB法不能提取出檀香心材总RNA,而SDS酸酚法、改良CTAB法和异硫氰酸胍-SDS法均能提取檀香心材总RNA。SDS酸酚法的A260 nm/A230 nm小于2.0,且RNA产率低,仅为(27.94±1.06)μg g–1,不能满足后续实验要求。而改良CTAB法和异硫氰酸胍-SDS法提取的总RNA带型清晰,完整性好,A260 nm/A280 nm为1.8~2.0,A260 nm/A230 nm大于2.0,RNA产率分别为(79.06±4.22)和(107.00±1.36)μg g–1。分别以改良CTAB法和异硫氰酸胍-SDS法提取的总RNA为模板,通过RT-PCR反应,扩增檀香Actin基因片段,结果二者扩增产物大小相同且条带单一,说明改良CTAB法与异硫氰酸胍-SDS法为檀香心材总RNA提取的较好方法。In order to screen RNA extracting method from heartwood of Santalum album, five methods were compared, including Trizol method, improved CTAB method, SDS-phenol method, guanidine isothiocyanate-CTAB method, and guanidine isothiocyanate-SDS method. The results showed that it was difficult to extract totalRNA by Trizol method and guanidine isothiocyanate-CTAB method, and the improved CTAB method,SDS-phenol method and guanidine isothiocyanate-SDS method could successfully extract total RNA. TheSDS-phenol method was not suitable for RNA extraction, because A260 nm/A230 nm was less than 2.0 and total RNAyield was low with (27.94±1.06) μg g–1. However, the 28S rRNA and 18S rRNA bands of total RNA extracted byimproved CTAB method and guanidine isothiocyanate-SDS method were clear, A260 nm/A280 nm ranged from 1.8 to2.0, A260 nm/A230 nm were more than 2.0, the yields were (79.06±4.22) and (107.00±1.36) μg g–1, respectively. The Actin segments in total RNA extracted by improved CTAB method and guanidine isothiocyanate-SDS method could amplificated with the same size and sigle band. Therefore, the improved CTAB methodand and guanidine isothiocyanate-SDS method were effective methods to extract total RNA from heartwood of S. album.

关 键 词：檀香 心材 RNA 改良CTAB法 异硫氰酸胍-SDS法 

分 类 号：S792.99[农业科学—林木遗传育种]