RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction

被引：451

作者：

Gasic K. ^{[1
]}

Hernandez A. ^{[2
]}

Korban S.S. ^{[1
]}

机构：

[1] Department of Natural Resources and Environmental Sciences, University of Illinois, 310 Madigan Building, Urbana, IL 6180

[2] Biotechnology Keck Center, University of Illinois, 354 Madigan Building, Urbana, IL 61801

来源：

Plant Molecular Biology Reporter | 2004年 / 22卷 / 4期

关键词：

Apple; Bud; cDNA libraries; Flower; Fruit; RNA extraction; Shoot;

D O I：

10.1007/BF02772687

中图分类号：

学科分类号：

摘要：

Recovering RNA of high quality and quantity is a prerequisite for ensuring representation of all expressed genes in a cDNA library. An efficient procedure for isolating RNA from bud, internodal shoot, flower, and fruit tissues of apple has been developed. This protocol does not involve the use of phenol, lyophilization, or ultracentrifugation. In addition, this protocol overcomes problems of both RNA degradation and low yield attributed to oxidation by polyphenolic compounds and coprecipitation with polysaccharides, both abundant components in apple fruit and flower tissues. Isolated RNA is of high quality and is undegraded as assessed by spectrophotometric readings and electrophoresis in denaturing agarose gels. RNA quality is further assessed following its use in reverse transcription and cDNA library construction, and it can be used for a number of downstream analyses, including Northern blot hybridization and reverse transcription-polymerase chain reaction (RT-PCR). With this modified protocol, 25-900 μg of total RNA is routinely obtained from 1 g of fresh material. This method is of low cost and easy to perform. © 2004 International Society for Plant Molecular Biology.

引用

页码：437 / 438

页数：1

共 11 条

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