【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。更多还原

【Abstract】 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 isothiocyanateCTAB method, and guanidine isothiocyanate-SDS method. The results showed that it was difficult to extract total RNA 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. The SDS-phenol method was not suitable for RNA extraction, because A260 nm/ A230 nm was less than 2.0 and total RNA yield was low with(27.94 ± 1.06) μg g–1. However, the 28 S r RNA and 18 S r RNA bands of total RNA extracted by improved CTAB method and guanidine isothiocyanate-SDS method were clear, A260 nm/ A280 nm ranged from 1.8 to 2.0, 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.更多还原