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Effective and economical column-based method for RNA isolation from animal cells

Biotechnology Letters volume 41, pages 915–920 (2019)Cite this article

Abstract

Objectives

To develop an efficient, economical, and low-toxicity method for the extraction of RNA from animal cells to meet a basic requirement of biological research: the isolation of high-quality RNA.

Results

Guanidine hydrochloride was used as a lysis buffer and Na-acetate was used as a wash buffer to extract RNA fragments from TM3 Leydig cells and ovarian granulosa cells efficiently. The functionality of the extracted RNA samples was verified through polymerase chain reaction (PCR) and real-time fluorescence quantitative PCR (RT-PCR). PCR results showed that the normal DNA column-based method could guarantee RNA integrity and could be used to amplify gene fragments successfully. RT-PCR analysis showed that the RNA samples isolated through the proposed method could be used to detect the expression levels of steroidogenic acute regulatory protein and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 mRNA in TM3 Leydig cells under induction by luteinizing hormone. The proposed method could be used to isolate RNA from mammalian cells and provided RNA yields of > 120 ng/5 × 106 cells. This method provided RNA with purities and yields that are sufficient for cDNA synthesis and PCR amplification in gene expression studies.

Conclusions

The proposed RNA extraction method has the advantages of low toxicity, safe handling, and low cost. Isolation can be completed in 20 min. The proposed method can be used to extract RNA from various animal cell samples and is worth promoting.

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Acknowledgements

This work was supported by The National Natural Science Foundation of China (Grant Nos. 31672417 and 31301969), Jilin Key Programs for Science and Technology Development (Grant No. 20170204039NY).

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Author notes

  1. Qiaoge Niu and Maosheng Cao are the Co-first authors.

Affiliations

  1. College of Animal Sciences, Jilin University, 5333 Xian Road, Changchun, 130062, Jilin, China

    Qiaoge Niu, Maosheng Cao, Shuxiong Chen, Yun Zhao, Lu Chen, Chunjin Li & Xu Zhou

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  1. Qiaoge Niu
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  2. Maosheng Cao
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  3. Shuxiong Chen
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  4. Yun Zhao
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  5. Lu Chen
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  6. Chunjin Li
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  7. Xu Zhou
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Correspondence to Chunjin Li or Xu Zhou.

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Niu, Q., Cao, M., Chen, S. et al. Effective and economical column-based method for RNA isolation from animal cells. Biotechnol Lett 41, 915–920 (2019). https://doi.org/10.1007/s10529-019-02700-7

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  • DOI: https://doi.org/10.1007/s10529-019-02700-7

Keywords

  • Animal cells
  • Column
  • Ethanol
  • Guanidine hydrochloride
  • RNA isolation