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[1]罗绍银,高继海,叶生亮,等.一种提取富含油脂植物组织总RNA的改良SDS酸酚法[J].应用与环境生物学报,2009,15(02):284-287.[doi:10.3724/SP.J.1145.2009.00284]
 LUO Shaoyin,GAO Jihai,YE Shengliang,et al.An Improved SDS Acid Phenol Method for Total RNA Extraction from Plant Tissues Rich in Lipid[J].Chinese Journal of Applied & Environmental Biology,2009,15(02):284-287.[doi:10.3724/SP.J.1145.2009.00284]
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一种提取富含油脂植物组织总RNA的改良SDS酸酚法()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
15卷
期数:
2009年02期
页码:
284-287
栏目:
生物质能源专辑
出版日期:
2009-03-15

文章信息/Info

Title:
An Improved SDS Acid Phenol Method for Total RNA Extraction from Plant Tissues Rich in Lipid
作者:
罗绍银高继海叶生亮徐莺陈放
(四川大学生命科学学院生物资源与生态环境教育部重点实验室 成都 610064)
Author(s):
LUO Shaoyin GAO Jihai YE Shengliang XU Ying** & CHEN Fang
(Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, School of Life Sciences, Sichuan University, Chengdu 610064, China)
关键词:
总RNA麻疯树油脂SDS酸酚法胚乳
Keywords:
total RNA Jatropha curcas oil SDS acid phenol method endosperm
分类号:
Q949.753.506
DOI:
10.3724/SP.J.1145.2009.00284
文献标志码:
A
摘要:
采用常规的方法提取富含油脂的植物组织总RNA常难以取得满意的结果. 对林莎两次裂解法进行了改良,即将SDS的浓度降低至1%,所用酸酚的pH值降低至4.0,去掉提取液II处理步骤,获得的总RNA效果较好:得率高,RNA产量可达74.7 μg (100 mg)-1;没有DNA污染,A260 nm/A280 nm值为2.026,A260 nm/A230 nm值为2.082,说明蛋白质及其它污染非常低,通过RT-PCR成功地扩增出了麻疯树curcin基因长为927 bp的特异条带. 相比林莎的方法,改良SDS酸酚法具有经济、简便、适用性广的特点. 图5 表1 参13
Abstract:
The total RNA extraction from plant tissues richin lipid by conventional methods often cannot meet the requirement of downstream experiments. This problem was overcomed by modifying Lin’s twice-extraction method: Adjusting the concentration of SDS and pH value of phenol to 1% and 4.0 respectively, and removing the solution II treatment. The yield was high and reached 74.7 μg (100 mg)-1 (fresh weight). There was no genomic DNA contamination in the process. The value of A260 nm/A280 nm was 2.026 and that of A260 nm/A230 nm was 2.082, meaning that protein and other contaminations were low. A 927 bp fragment of Jatropha curcas’ curcin gene was successfully amplified from the obtained total RNA. Compared to Lin’s method, this improved SDS acid phenol method is economical, convenient and has a wide applicability. Fig 5, Tab 1, Ref 13

参考文献/References:

1 Lin S (林莎), Luo SY (罗绍银), Lin Y (林颖), Wang ZT (王治涛), Niu P (牛蓓), Qin XB (秦小波), Li YQ (李月琴), Xu Y (徐莺), Chen F (陈放). A simple method for extracting total RNAs from endosperm of Jatropha curcas L. Chin J Appl Environ Biol (应用与环境生物学报), 2008, 14 (5): 692~694
2 Bμgos RC, Chiang VL, Zhang XH, Campbell ER, Podila GK, Campbell WH. RNA isolation from plant tissues recalcitrant to extraction in guanidine. Biol Techniques, 1992, 19: 734~737
3 Schneiderbauer A, Sandermann H, Jr Ernst D. Isolation of functional RNA from plant tissues rich in phenolic compounds. Anal Biochem, 1991, 197: 91~95
4 Groppe JC, Morse DE. Isolation of full-length RNA templates for reverse transcription from tissues rich in RNase and proteoglycans. Anal Biochem, 1993, 210: 337~343
5 Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonucleases. Biochemistry, 1979, 18: 5294~5299
6 Gesteira Ada S, Micheli F, Ferreira CF, Cascardo JC. Isolation and purification of functional total RNA from different organs of cacao tree during its interaction with the pathogen Crinipillis perniciosa. Biol Techniques, 2003, 35: 495~499
7 Murillo I, Raventos D, Jaeck E, San Segundo B. Isolation of total RNA and mRNA from plant tissues. Promega Notes Mag, 1995, 54: 2~7
8 Li H (李化), Chen L (陈丽), Tang L (唐琳), Chen F (陈放). Physicochemical characteristics and fatty-acid composition of seed oil of Jatropha curcas from Southwest China. Chin J Appl Environ Biol (应用与环境生物学报), 2006, 12 (5): 643~646
9 ABE S, Fujisawa T, Satake M, Gata K. Studies on SDS-phenol methods for extraction of rat liver nuclear RNA I. Purity, recovery, and specific radioactivity of pulse labeled nuclear RNA obtained by SDS-phenol extraction under various condition. J Biochem, 1972, 72: 561~570
10 ABE S, Fujisawa T, Satake M, Gata K. Studies on SDS-phenol methods for extraction of rat liver nuclear RNA II. Polyacrylamide gel electrophoresis of nuclear RNA obtained using various conditions for SDS-phenol extraction. J Biochem, 1972, 72: 571~581
11 Wang YH (王友华), Lu MZ (卢孟柱), Duan LS (段留生). Modified hot phenol method for extracting total RNA from seedling root of Cotton. Acta Bot Bor-Occid Sin (西北植物学报), 2005, 25 (4): 723~726
12 Smbrook J, Russell DW. Molecular cloning. 3th ed. Huang PT (黄培堂). Cold Spring Harbor Laboratory Press, 2002. 518
13 Chomzynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate -phenol-chloroform extraction. Anal Biochem, 1987, 162: 156~159

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备注/Memo

备注/Memo:
国家自然科学基金项目(No. 30670204)、科技部国际合作项目(No. 2006DFB63400)、教育部博士点基金项目(No. 20060610015)和国家“十一五”科技支撑项目(No. 2006BAD07A04)资助 Supported by the National Natural Science Foundation of China (No. 30670204), the International Cooperation Program of Ministry of Sci & Tech of China (No. 2006DFB63400), the Ph. D. Program Foundation of Ministry of Education of China (No. 20060610015) and the “11th Five-year-plan” Major Program of China (No. 2006BAD07A04)
更新日期/Last Update: 2009-05-05