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使用电荷耦合器件(CCD)相机成像系统作为记录设备,通过狭缝印迹杂交对人类DNA进行定量分析。

Using a CCD camera imaging system as a recording device to quantify human DNA by slot blot hybridization.

作者信息

Budowle B, Hudlow W R, Lee S B, Klevan L

机构信息

FBI Academy, Quantico, VA, USA.

出版信息

Biotechniques. 2001 Mar;30(3):680-5. doi: 10.2144/01303pf01.

Abstract

Slot blot hybridization of membrane-immobilized, single-stranded human DNA with the higher primate-specific alphoid probe D17Z1 is routinely used in forensic science to estimate the amount of DNA in biological samples. Typically, a chemiluminescent signal captured on film records the hybridization, and the quantity of the signal is related to the amount of immobilized DNA. Digital imaging using a cooled CCD camera offers an alternate non-film-based method for image acquisition with comparable sensitivity of detection, a greater dynamic range, enhanced capability of data interpretation, and often faster results than film. In addition, the data support the premise that more accurate and precise human DNA quantification should be obtained by not assuming a linear response of signal to known standards. Instead, quantity should be estimated using a second-order standard curve (R2 = 0.999). Finally, a CCD camera imaging system offers versatility for image capture of different signal sources and analysis of samples on a variety of support media.

摘要

利用高等灵长类特异性α卫星探针D17Z1对膜固定的单链人类DNA进行狭缝印迹杂交,在法医学中常用于估计生物样本中的DNA含量。通常,胶片上捕获的化学发光信号记录杂交情况,信号量与固定的DNA量相关。使用冷却电荷耦合器件(CCD)相机的数字成像提供了一种基于非胶片的替代图像采集方法,具有可比的检测灵敏度、更大的动态范围、更强的数据解读能力,并且通常比胶片能更快得到结果。此外,数据支持这样一个前提,即不应假设信号对已知标准呈线性响应,而应通过二阶标准曲线(R2 = 0.999)来估计含量,从而获得更准确和精确的人类DNA定量。最后,CCD相机成像系统具有通用性,可用于捕获不同信号源的图像,并分析各种支持介质上的样本。

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