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Effects of Delay in the Snap Freezing of Colorectal Cancer Tissues on the Quality of DNA and RNA

대한대장항문학회지 2010년 26권 5호 p.316 ~ 323
 ( Hong Sang-Hyun ) - 전북대학교 의과대학 병리학교실

 ( Baek Hyun-Ah ) - 전북대학교 의과대학 병리학교실
장규윤 ( Jang Kyu-Yun ) - 전북대학교 의과대학 병리학교실
정명자 ( Chung Myoung-Ja ) - 전북대학교 의과대학 병리학교실
문우성 ( Moon Woo-Sung ) - 전북대학교 의과대학 병리학교실
강명재 ( Kang Myoung-Jae ) - 전북대학교 의과대학 병리학교실
이동근 ( Lee Dong-Geun ) - 전북대학교 의과대학 병리학교실
박호성 ( Park Ho-Sung ) - 전북대학교 의과대학 병리학교실

Abstract


Purpose The success of basic molecular research using biospecimens strongly depends on the quality of the specimen. In this study, we evaluated the effects of delayed freezing time on the stability of DNA and RNA in fresh frozen tissue from patients with colorectal cancer.

Methods Tissues were frozen at 10, 30, 60, and 90 minutes after extirpation of colorectal cancer in 20 cases. Absorbance ratio of 260 to 280 nm (A260/A280) and agarose gel electrophoresis were evaluated. In addition, the RNA integrity number (RIN) was assayed for the analysis of the RNA integrity.

Results Regardless of delayed freezing time, all DNA and RNA samples revealed A260/A280 ratios of more than 1.9, and all DNA samples showed a discrete, high-molecular-weight band on agarose gel electrophoresis. The RINs were 7.53 ± 2.04, 6.70 ± 1.88, 6.47 ± 2.58, and 4.22 ± 2.34 at 10, 30, 60, and 90 minutes, respectively. Though the concentration of RNA was not affected by delayed freezing, the RNA integrity was decreased with increasing delayed freezing time.

Conclusion According to the RIN results, we recommend that the collection of colorectal cancer tissue should be done within 10 minutes for studies requiring RNA of high quality and within 30 minutes for usual RNA studies.

키워드

Colorectal neoplasms; Tissue banks; DNA; RNA; Quality control
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