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A Modified Method using TRIzol® Reagent and Liquid Nitrogen Produces High-Quality RNA from Rat Pancreas

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Abstract

To establish an economical and reproducible method for the high-quality RNA extraction from pancreas, we isolated total RNA from rat pancreas with TRIzol® reagent and liquid nitrogen. In the initial stage, we optimized three influential factors, the way to homogenize pancreas, the time to collect the pancreatic tissue from animals, and the weight of the pancreatic tissue in 1 ml of TRIzol® reagent. The RNA quality was determined by detecting total RNA content and its absorbance at 260/280 nm wavelength, visualizing RNA in non-denatured agarose gel and performing RT-PCR of pancreas-specific genes. The A 260/A 280 ratio of the total RNA extracted by grinding 20–30 mg of rat pancreatic tissue removed from the rats in liquid nitrogen within 1 min and then immersed in 1 ml of the TRIzol® Reagent was 1.75–1.89, and the ratio of 28S/18S ribosomal RNA bands was more than 1.8. Furthermore, full length of Pdx1 open-reading frame was amplified with RNA extracted from the grinding group rather than from the conventional group. The RT-PCR products of pancreas-specific genes from both exocrine and endocrine parts of pancreas were successfully derived from the extracted RNA. The results suggested that we successfully provided an economical, fast, and reproducible method to obtain the high-quality and intact RNA from rat pancreas with TRIzol® Reagent and liquid nitrogen.

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Acknowledgments

We are very grateful to Dr. Hongmin Li (School of Life Science, Northwest University, Xi’an, China) for her excellent technical assistance and to Mr. Muhanmude Shahzad for critical reading and linguistic correction of the manuscript.

Funding

The project was supported by the National Natural Science Foundation of China (No. 30400249, 30571725 and 30630058), the Shaanxi Province International Cooperational Foundation of China (No.2007-kw-06), and the Shaanxi Province Natural Science Foundation of China (No. 2004C256).

Conflict of interest

The authors declare no competing interests.

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Authors and Affiliations

  1. Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education of China, Xi’an Jiaotong University School of Medicine, Xi’an, Shaanxi, 710061, People’s Republic of China

    Dongmin Li, Wuchao Ren, Xuan Wang, Feimiao Wang, Yu Gao, Qilan Ning, Yan Han, Tianbao Song & Shemin Lu

  2. Department of Genetics and Molecular Biology, Xi’an Jiaotong University School of Medicine, Xi’an, Shaanxi, 710061, People’s Republic of China

    Dongmin Li, Wuchao Ren, Xuan Wang, Feimiao Wang, Yu Gao, Qilan Ning, Yan Han & Shemin Lu

  3. Department of Human Anatomy, Histology and Embryology, Xi’an Jiaotong University School of Medicine, Xi’an, Shaanxi, 710061, People’s Republic of China

    Tianbao Song

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  1. Dongmin Li
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  2. Wuchao Ren
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  3. Xuan Wang
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  4. Feimiao Wang
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  5. Yu Gao
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  6. Qilan Ning
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  7. Yan Han
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  8. Tianbao Song
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  9. Shemin Lu
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Corresponding author

Correspondence to Dongmin Li.

Additional information

Dongmin Li and Wuchao Ren equally contributed to this work. Shemin Lu and Tianbao Song are co-corresponding author.

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Li, D., Ren, W., Wang, X. et al. A Modified Method using TRIzol® Reagent and Liquid Nitrogen Produces High-Quality RNA from Rat Pancreas. Appl Biochem Biotechnol 158, 253–261 (2009). https://doi.org/10.1007/s12010-008-8391-0

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  • DOI: https://doi.org/10.1007/s12010-008-8391-0

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