Laboratório de Genética Aplicada, Instituto de Estudos Costeiros, Universidade Federal do Pará, Bragança, Pará, Brazil.
Laboratório de Genética e Biologia Molecular, Instituto de Estudos Costeiros, Universidade Federal do Pará, Bragança, Pará, Brazil.
PLoS One. 2023 Feb 28;18(2):e0282369. doi: 10.1371/journal.pone.0282369. eCollection 2023.
Molecular genetic techniques are an effective monitoring tool, but high-quality DNA samples are usually required. In this study, we compared three different protocols of DNA extraction: NaCl (saline); phenol-chloroform and commercial kit (Promega)-from three biological tissues of five individuals of Lutjanus purpureus under two methods of storage. The evaluated items included DNA concentration and purity, processing time and cost, as well as the obtaining of functional sequences. The highest average values of DNA concentration were obtained using the saline procedure and the commercial kit. Pure DNA was only obtained using the saline protocol, evaluated by the ratio of 260/280. The saline and phenol-chloroform protocols were the least expensive methods. The commercial kit costs are counterbalanced by the short time required. The procedure based on phenol-chloroform presented the worst results regarding DNA yield and the time required to perform all steps. The saline and commercial kit protocols showed similar results concerning the amount and quality of extracted DNA. Therefore, the final choice should be based on the available financial resources and the available time for carrying out each procedure of DNA extraction.
分子遗传学技术是一种有效的监测工具，但通常需要高质量的 DNA 样本。在这项研究中，我们比较了三种不同的 DNA 提取方案：NaCl（生理盐水）；苯酚-氯仿法和商业试剂盒（Promega）-从 5 个紫红笛鲷个体的三种生物组织中，采用两种存储方法。评估的项目包括 DNA 浓度和纯度、处理时间和成本，以及获得功能序列。使用生理盐水方案和商业试剂盒可获得最高的 DNA 浓度平均值。仅使用生理盐水方案可获得纯 DNA，通过 260/280 的比值进行评估。生理盐水和苯酚-氯仿法是最便宜的方法。商业试剂盒的成本可以通过所需的短时间来平衡。基于苯酚-氯仿的方法在 DNA 产量和执行所有步骤所需的时间方面表现最差。生理盐水和商业试剂盒方案在提取的 DNA 量和质量方面表现相似。因此，最终的选择应基于可用的财务资源和执行每个 DNA 提取步骤的可用时间。
