A convenient and efficient protocol for isolating high-quality RNA from latex of Hevea brasiliensis (para rubber tree)
文献类型: 外文期刊
作者: Tang, Chaorong 1 ; Qi, Jiyan 1 ; Li, Heping 1 ; Zhang, Cunliang 1 ; Wang, Yuekun 1 ;
作者机构: 1.Chinese Acad Trop Agr Sci, Rubber Res Inst, Key Lab Minist Agr Trop Crops Physiol, Danzhou 571737, Peoples R China
关键词: Hevea brasiliensis;latex;RNA isolation;RNA quality;liquid nitrogen;ultra-low temperature
期刊名称:JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS ( 影响因子:1.808; 五年影响因子:2.048 )
ISSN: 0165-022X
年卷期: 2007 年 70 卷 5 期
页码:
收录情况: SCI
摘要: isolating high-quality RNA from latex of H. brasiliensis is a prerequisite to elucidating the molecular mechanisms of rubber biosynthesis and its regulation. Here, an improved protocol was developed for latex collection, transportation, storage, and RNA isolation. Compared with existing ones, our protocol eliminated liquid nitrogen for latex collection and subsequent low-temperature (-70 degrees C) condition for latex storage, making it more convenient and feasible when latex was collected in remote sampling sites, and latex storage and RNA isolation were conducted in poorly-equipped laboratories. Different methods (UV absorbance scans, denaturing gel electrophoresis, autoradiograph monitoring of cDNA synthesis) were used to confirm the high quality of the RNA prepared with this protocol, whose usefulness was further verified by several practical applications, including construction of one high-quality cDNA library, cloning of the full-length cDNAs of 3 novel Hevea sucrose transporter genes, and semi-quantitative RT-PCR analysis of two rubber-biosynthesis essential genes and one sucrose transporter gene. (c) 2007 Elsevier B.V. All rights reserved.
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