AN EFFECTIVE METHOD OF RNA ISOLATION FROM Fallopia multiflora TUBEROUS ROOTS

被引:6
作者
Chen, Lei [1 ]
Sheng, Shu-Jing [2 ,3 ]
Tan, Xue-mei [2 ]
Shen, Yan-Jing [2 ]
Li, Hong-qing [1 ]
Zhao, Shu-jin [2 ]
机构
[1] S China Normal Univ, Guangdong Prov Key Lab Biotechnol Plant Dev, Coll Life Sci, Guangzhou, Guangdong, Peoples R China
[2] Gen Hosp Guangzhou Mil Command, Dept Pharm, Guangzhou, Guangdong, Peoples R China
[3] S China Univ Technol, Sch Biosci & Bioengn, Guangzhou, Guangdong, Peoples R China
关键词
cDNA library; Fallopia multiflora; Northern blot; RNA isolation; RT-PCR; tuberous roots; HIGH-QUALITY RNA; RICH; POLYPHENOLS; EXTRACTION;
D O I
10.1080/10826068.2011.566297
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To isolate high-quality total RNA from Fallopia multiflora tuberous roots is difficult because of the presence of high levels of carbohydrates, phenolics, and other secondary metabolites. Since several procedures specialized for RNA isolation from polysaccharides and phenols rich tissues have resulted in poor yields, in this study, we developed a modified protocol that was derived from the traditional CTAB method. The protocol was able to produce high-quality and intact RNA from the tuberous roots of F. multiflora. The yield of total RNA was more than 0.15 mg/g fresh weight, with an A260/A280 ratio of 1.9-2.0. The obtained RNA was of sufficient quality and suitable for downstream application such as reverse-transcription polymerase chain reaction (RT-PCR), Northern hybridization, and cDNA library construction. The protocol may also have wider applicability for total RNA isolation from other plant species with tuberous roots.
引用
收藏
页码:87 / 96
页数:10
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