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Recovery of MicroRNA from Stored Human Peripheral Blood Samples

机译:从人类外周血样本中回收MicroRNA

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Recovery of microRNAs (miRs) from tissue samples of many kinds has become an area of increasing interest and importance in recent years. We isolated and then amplified and quantitated miRs from human peripheral blood samples stored in the University of Colorado Denver Skin Cancer Biorepository to determine whether miR recovery was possible and consistent over time in storage. Forty-five blood samples from patients with different stages of malignant melanoma were collected in PAXgene Blood RNA tubes and then stored at -80 ℃ prior to RNA preparation. The samples examined had been stored from 4 weeks to 3 years. Total RNA was prepared, followed by miR isolation, amplification, and quantitation using real-time polymerase chain reaction. A widely expressed miR, miR-221, was used as a standard for comparison across samples and storage time. miR-221 was recovered from all samples, with no differences observed with longer storage time. These studies show that miRs can be recovered and quantified from human blood samples stored for up to 3 years.
机译:近年来,从多种组织样品中回收microRNA(miR)成为人们越来越感兴趣和重视的领域。我们从科罗拉多大学丹佛分校皮肤癌生物储存库中储存的人类外周血样本中分离出miRs,然后对其进行了扩增和定量分析,以确定miR回收是否可能且在储存过程中保持一致。在PAXgene血液RNA管中收集了来自不同阶段恶性黑色素瘤患者的45份血液样本,然后在制备RNA之前保存在-80℃。检查的样品已保存4周至3年。制备总RNA,然后使用实时聚合酶链反应进行miR分离,扩增和定量。广泛表达的miR,miR-221被用作比较样品和保存时间的标准。从所有样品中回收了miR-221,较长的存储时间未发现差异。这些研究表明,miRs可以从长达3年的人类血液样本中回收和定量。

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