Method Optimization for Extracting High-Quality RNA From the Human Pancreas Tissue.
Method Optimization for Extracting High-Quality RNA From the Human Pancreas Tissue.
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DOI：
10.1016/j.tranon.2018.04.004
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发表时间：
2018-06
影响因子：
5
通讯作者：
Kim SC
中科院分区：
文献类型：
--
作者：
Jun E;Oh J;Lee S;Jun HR;Seo EH;Jang JY;Kim SC
核酸测序经常用于确定疾病的分子基础。因此，生物样本的适当储存对于抑制核酸降解至关重要。从人胰腺分离的RNA通常质量差，因为其内源性RNA酶浓度高。在这项研究中，我们优化了从八名胰腺癌患者术后获得的配对肿瘤和正常胰腺组织中提取高质量RNA的方法。检查RNA完整性值（RIN）以评估RNA的完整性，我们尝试提取RIN值为8或更高的RNA，以便进行最新的遗传分析。检查了几个参数对提取的RNA的数量和质量的影响，包括用于组织裂解的方法、RNAlater处理、储存时的组织重量和手术切除后的储存时间。数据显示，使用组织裂解的手动和机械方法的组合分离最高量的RNA。此外，将组织切成小块（<100 mg）并在储存前用RNAlater溶液处理它们增加了RNA稳定性。根据这些指导原则，从100%（8/8）的肿瘤组织和75%（6/8）的正常组织中获得高质量的RNA。高质量的RNA在反复冻融条件下仍保持稳定。这些结果在临床环境中的样本处理和储存期间的应用将促进疾病的遗传诊断及其随后的治疗。
Nucleic acid sequencing is frequently used to determine the molecular basis of diseases. Therefore, proper storage of biological specimens is essential to inhibit nucleic acid degradation. RNA isolated from the human pancreas is generally of poor quality because of its high concentration of endogenous RNase. In this study, we optimized the method for extracting high quality RNA from paired tumor and normal pancreatic tissues obtained from eight pancreatic cancer patients post-surgery. RNA integrity number (RIN) was checked to evaluate the integrity of RNA, we tried to extract the RNA with an RIN value of 8 or higher that allows for the latest genetic analysis. The effect of several parameters, including the method used for tissue lysis, RNAlater treatment, tissue weight at storage, and the time to storage after surgical resection, on the quantity and quality of RNA extracted was examined. Data showed that the highest quantity of RNA was isolated using a combination of manual and mechanical methods of tissue lysis. Additionally, sectioning the tissues into small pieces (<100 mg) and treating them with RNAlater solution prior to storage increased RNA stability. Following these guidelines, high quality RNA was obtained from 100% (8/8) of tumor tissues and 75% (6/8) of normal tissues. High-quality RNA was still stable under repeated freezing and thawing. The application of these results during sample handling and storage in clinical settings will facilitate the genetic diagnosis of diseases and their subsequent treatment.
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影响因子：
--
作者：
Schroeder A;Mueller O;Stocker S;Salowsky R;Leiber M;Gassmann M;Lightfoot S;Menzel W;Granzow M;Ragg T
通讯作者：
Ragg T
影响因子：
3.7
作者：
Rudloff U;Bhanot U;Gerald W;Klimstra DS;Jarnagin WR;Brennan MF;Allen PJ
通讯作者：
Allen PJ
DOI：
10.3393/jksc.2010.26.5.316
发表时间：
2010-10
期刊：
Journal of the Korean Society of Coloproctology
影响因子：
--
作者：
Hong SH;Baek HA;Jang KY;Chung MJ;Moon WS;Kang MJ;Lee DG;Park HS
通讯作者：
Park HS
影响因子：
14.9
作者：
Imbeaud S;Graudens E;Boulanger V;Barlet X;Zaborski P;Eveno E;Mueller O;Schroeder A;Auffray C
通讯作者：
Auffray C
影响因子：
1.6
作者：
Condelli, Valentina;Lettini, Giacomo;Landriscina, Matteo
通讯作者：
Landriscina, Matteo