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A Modified Protocol for RNA Extraction from Different Peach Tissues Suitable for Gene Isolation and Real-Time PCR Analysis

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Abstract

RNA extraction is the first step in the study of gene isolation and expression. However, it is difficult to extract high quantity and quality RNA from tissues containing large quantities of polysaccharides and polyphenols. Peach (Prunus persica), in addition to containing high levels of polysaccharides and polyphenols, is a challenging starting material for RNA isolation using a single method because of different amounts of those substances in diverse tissues. Based on three reported methods, we developed a modified RNA isolation protocol to solve this problem, leading to high quality and quantity of total RNA from peach mesocarp tissues of fruits which were sampled from all developmental stages and different storage periods, as well as from other tissues including flowers, leaves, stems, and roots. With our modified method, 28–650 μg of total RNA was routinely obtained from per gram of fresh material, gave at least a 1.16-fold improvement by compared with those isolated by other seven methods. The RNA extracts were successfully used in downstream applications such as RT-PCR, RACE, and real-time PCR.

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Acknowledgments

The authors gratefully acknowledge the financial support extended by the National Natural Science Foundation of China (30370987; 30871681) and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) for this study. The authors also greatly appreciate the critical review of the manuscript by Dr. Zhen-Xiang Lu, Lethbridge Research Centre, Agriculture and Agri-Food Canada, Lethbridge, Alberta, Canada.

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Authors and Affiliations

  1. College of Horticulture, Nanjing Agricultural University, 1 Tongwei Road, Weigang, Nanjing, 210095, Jiangsu, People’s Republic of China

    Zhaoguo Tong, Shenchun Qu, Jiyu Zhang, Fei Wang, Jianmin Tao, Zhihong Gao & Zhen Zhang

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  1. Zhaoguo Tong
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  2. Shenchun Qu
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  3. Jiyu Zhang
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  4. Fei Wang
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  5. Jianmin Tao
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  6. Zhihong Gao
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  7. Zhen Zhang
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Corresponding authors

Correspondence to Zhihong Gao or Zhen Zhang.

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Z. Tong and S. Qu contributed equally to this work.

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Tong, Z., Qu, S., Zhang, J. et al. A Modified Protocol for RNA Extraction from Different Peach Tissues Suitable for Gene Isolation and Real-Time PCR Analysis. Mol Biotechnol 50, 229–236 (2012). https://doi.org/10.1007/s12033-011-9433-3

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