A novel approach to imaging and visualization of minute amounts of DNA in small volume samples,Analyst

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A novel approach to imaging and visualization of minute amounts of DNA in small volume samples
Analyst ( IF 3.6 ) Pub Date : 2021-09-20 , DOI: 10.1039/d1an01391b
Luca Ceresa ₁ , Emma Kitchner ₁ , Michael Seung ₁ , Magdalena M Bus _{2,

3} , Bruce Budowle _{2,

3} , Jose Chavez ₁ , Ignacy Gryczynski ₁ , Zygmunt Gryczynski ₁

Affiliation

This report presents a novel approach for detecting and visualizing small to trace amounts of DNA in a sample. By utilizing both the change in emission spectrum and change in fluorescence lifetime, there is a significant increase in detection sensitivity allowing for the imaging/visualizing of a picograms amount of DNA in a microliters volume. As in the previous reports, one of the oldest DNA intercalators, Ethidium Bromide (EtBr), is employed as a model system. With this new approach, it is feasible to visualize just a few hundred picograms of DNA without the need for prior DNA amplification. The sensitivity can later be largely improved by using an intercalator that exhibits a higher affinity to DNA and a larger fluorescence change upon binding to DNA (e.g., ethidium homodimer, YOYO, or Diamond nucleic acid dyes).

中文翻译：

一种在小体积样品中对微量 DNA 进行成像和可视化的新方法

本报告提出了一种检测和可视化样本中少量到痕量 DNA 的新方法。通过利用发射光谱的变化和荧光寿命的变化，检测灵敏度显着提高，允许对微升体积中皮克量的 DNA 进行成像/可视化。与之前的报告一样，最古老的 DNA 嵌入剂之一溴化乙锭 (EtBr) 被用作模型系统。使用这种新方法，无需事先进行 DNA 扩增，只需可视化几百皮克的 DNA 是可行的。稍后可以通过使用对 DNA 具有更高亲和力和与 DNA 结合时荧光变化更大的嵌入剂来大大提高灵敏度（例如、乙锭同二聚体、YOYO 或钻石核酸染料）。

更新日期：2021-09-24

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