从野外条件下保存的禽类有核全血中回收 RNA：商业方法的评估。

Recovery of RNA from avian nucleated whole blood stored under field conditions: an evaluation of commercial methods.

Department of Biological Applications and Technology, University of Ioannina, Ioannina, Greece.

Br Poult Sci. 2022 Oct;63(5):730-734. doi: 10.1080/00071668.2022.2061838. Epub 2022 Apr 20.

Abstract

Whole blood can be used to gain insights on several molecular mechanisms involved in animal physiology. However, its use to extract RNA from organisms that carry nucleated red blood cells is limited due to its high nuclease content and instability under field conditions. 2. Here, we evaluate three commercially available RNA preservation and extraction methods applied to avian whole-blood samples kept in different storage conditions to identify the most suitable ones for use in field studies. 3. Whole-blood starting volume was the most important factor; only samples with a starting volume of 50 µl yielded RNA, whereas all 100 µl samples failed to provide any RNA regardless of the method used. RNA concentration decreased with storage time in every method employed. Samples stored at -20°C yielded higher RNA concentration than the ones stored at 4°C, although some possible effects of freeze-thaw cycles were observed. 4. For two of the three methods, it is likely that DNA contamination and/or extensive degradation of RNA could have taken place. Only whole blood stored and extracted using the Quick RNA Whole Blood kit provided high-quality RNA in every condition tested.

摘要

全血可用于深入了解动物生理学中涉及的几种分子机制。然而，由于其核酸酶含量高且在野外条件下不稳定，其用于提取含有有核红细胞的生物体的 RNA 受到限制。
在这里，我们评估了三种市售的 RNA 保存和提取方法在不同储存条件下应用于禽类全血样本，以确定最适合野外研究的方法。
全血起始体积是最重要的因素；只有起始体积为 50µl 的样本才能产生 RNA，而无论使用哪种方法，所有 100µl 的样本均未能提供任何 RNA。在所使用的每种方法中，RNA 浓度随储存时间的延长而降低。在每种方法中，-20°C 储存的样本比 4°C 储存的样本产生更高浓度的 RNA，尽管观察到了一些冻融循环的可能影响。
对于其中两种方法，很可能发生了 DNA 污染和/或 RNA 的广泛降解。只有使用 Quick RNA Whole Blood 试剂盒储存和提取的全血在测试的每种条件下都提供了高质量的 RNA。