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High-quality total RNA extraction from Magnolia sieboldii K. Koch seeds: a comparative evaluation

Author:
Wei, Jun, Zhang, Xiaolin, Hou, Zhe, Lu, Xiujun
Source:
Journal of forestry research 2019 v.30 no.1 pp. 371-379
ISSN:
1007-662X
Subject:
Magnolia sieboldii, RNA, dormancy, germination, molecular biology, protocols, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, secondary metabolites, seeds, tissues, transcriptomics
Abstract:
Magnolia sieboldii K. Koch seed is characterized with having deep dormancy. The inner molecular regulation mechanism has not been investigated because of the absence of a protocol for total RNA (ribonucleic acid) extraction. The extraction of high-quality RNA is important and can be a limiting factor in plant molecular biology experiments. Sufficient high-quality total RNA is required to elucidate the molecular regulation mechanism of germination. However, M. sieboldii seeds with large amounts of secondary metabolites also contain recalcitrant tissues for RNA isolation. We found two simple and low-cost RNA extraction methods for M. sieboldii seeds by evaluating and selecting eight types of methods and further optimizing these methods. The two methods were not only suitable for extracting M. sieboldii seed RNA but also applicable to RNAs from several other tissues. Total RNA extracted through these approaches was applicable for general molecular biology experiments such as qRT-PCR (quantitative real-time polymerase chain reactions). The protocols also meet the strict harsh requirements for transcriptome sequencing and small RNA sequencing. This study provides a powerful approach for future studies at the transcription level.
Agid:
6275569