Buffered Lugol's Iodine Preserves DNA Fragment Lengths.

IF 2.2 4区 生物学 Q2 BIOLOGY
Integrative Organismal Biology Pub Date : 2024-06-14 eCollection Date: 2024-01-01 DOI:10.1093/iob/obae017
P M Gignac, D Valdez, A C Morhardt, L M Lynch
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Abstract

Museum collections play a pivotal role in the advancement of biological science by preserving phenotypic and genotypic history and variation. Recently, contrast-enhanced X-ray computed tomography (CT) has aided these advances by allowing improved visualization of internal soft tissues. However, vouchered specimens could be at risk if staining techniques are destructive. For instance, the pH of unbuffered Lugol's iodine (I2KI) may be low enough to damage deoxyribonucleic acid (DNA). The extent of this risk is unknown due to a lack of rigorous evaluation of DNA quality between control and experimental samples. Here, we used formalin-fixed mice to document DNA concentrations and fragment lengths in nonstained, ethanol-preserved controls and 3 iodine-based staining preparations: (1) 1.25% weight-by-volume (wt/vol.) alcoholic iodine (I2E); (2) 3.75% wt/vol. I2KI; and (3) 3.75% wt/vol. buffered I2KI. We tested a null hypothesis of no significant difference in DNA concentrations and fragment lengths between control and treatment samples. We found that DNA concentration decreases because of staining-potentially an effect of measuring intact double-stranded DNA only. Fragment lengths, however, were significantly higher for buffered I2KI and control samples, which were not, themselves, significantly different. Our results implicate buffered I2KI as the appropriate choice for contrast-enhanced CT imaging of museum wet collections to safely maximize their potential for understanding genetic and phenotypic diversity.

缓冲鲁戈尔碘保留 DNA 片段长度
博物馆藏品通过保存表型和基因型的历史和变异,在推动生物科学发展方面发挥着举足轻重的作用。最近,对比度增强型 X 射线计算机断层扫描(CT)改善了内部软组织的可视化,从而为这些进步提供了帮助。然而,如果染色技术具有破坏性,则凭证标本可能会面临风险。例如,未经缓冲的卢戈尔碘(I2KI)的 pH 值可能低到足以破坏脱氧核糖核酸(DNA)。由于缺乏对对照样本和实验样本 DNA 质量的严格评估,这种风险的程度尚不清楚。在此,我们使用福尔马林固定的小鼠记录未染色、乙醇保存的对照组和 3 种碘染色制剂中的 DNA 浓度和片段长度:(1) 1.25% 重量体积比(wt/vol. )的酒精碘 (I2E);(2) 3.75% 重量体积比(wt/vol. )的酒精碘 (I2KI);(3) 3.75% 重量体积比(wt/vol. )的酒精碘 (I2KI)。缓冲 I2KI。我们测试了一个零假设,即对照样本和处理样本之间的 DNA 浓度和片段长度没有显著差异。我们发现 DNA 浓度会因染色而降低,这可能是只测量完整双链 DNA 的影响。但是,缓冲 I2KI 和对照样本的片段长度明显更高,而两者本身并无明显差异。我们的研究结果表明,缓冲 I2KI 是对博物馆湿藏品进行对比增强 CT 成像的适当选择,可安全地最大限度地发挥其了解遗传和表型多样性的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
3.70
自引率
6.70%
发文量
48
审稿时长
20 weeks
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