title: Efficient isolation of high-quality RNA from lotus Nelumbo  nucifera ssp nucifera tissues.
abstract: Nelumbo nucifera is widely used as food, as an ornamental, in medicine,  and as packing material; it is also reported to have anti-HIV effects and  antioxidant capacity. We sought an improved method for extracting high-quality  total RNA from different tissues of N. nucifera. Four methods for RNA  extraction were assessed for their ability to recover high-quality RNA  applicable for evaluation of polyphenol oxidase (PPO) gene expression profiles.  The recovery and quality of the RNA obtained from five different tissues by the  best CTAB-LiCl method were evaluated through UV light absorbance. Both A(260)/A(280) and  A(260)/A(230) absorbance ratios were more than 2.0; the yield ranged  from 59.87 to 163.75 μg/g fresh weight. The brightness of the  28S band was approximately twice that of 18S; the latter was also considered as  high-quality RNA. The PPO gene fragment (606 bp) was successfully amplified by  RT-PCR, demonstrating the integrity of the isolated RNA. The relative  expression levels of the PPO gene based on RT-PCR in five tissues of lotus  were: rhizome buds (2.66), young leaves (2.42), fresh cut rhizome (2.02),  petals (1.80), and petiole (1.65), using housekeeping gene β-actin as an internal control. We  concluded that the total RNA isolated by this protocol is of sufficient quality  for molecular applications.
authors: Shaanxi Key Laboratory of Molecular Biology in Agriculture, College of Life Sciences, Northwest A & F University, Yangling, PR China.
authors: Y J
authors: YJ
authors: Zhang
authors: X Y
authors: XY
authors: Hao
authors: Z S
authors: ZS
authors: Liang
authors: W D
authors: WD
authors: Ke
authors: H B
authors: HB
authors: Guo
mesh_terms: D000199:Actins / Q000737:chemistry / Q000235:genetics; D004156:Catechol Oxidase / Q000235:genetics; D031653:Nelumbo / Q000737:chemistry* / Q000235:genetics*; D010936:Plant Extracts / Q000737:chemistry* / Q000235:genetics; D018749:RNA, Plant / Q000737:chemistry* / Q000235:genetics*
chemical_list: D000199:Actins; D010936:Plant Extracts; D018749:RNA, Plant; D004156:Catechol Oxidase