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用生物素化探针进行印迹杂交时检测亚皮克级特定DNA序列量。

Detection of sub-picogram quantities of specific DNA sequences on blot hybridization with biotinylated probes.

作者信息

Chan V T, Fleming K A, McGee J O

出版信息

Nucleic Acids Res. 1985 Nov 25;13(22):8083-91. doi: 10.1093/nar/13.22.8083.

DOI:10.1093/nar/13.22.8083
PMID:2933635
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC322111/
Abstract

A sensitive method for detecting biotinylated DNA probes on dot and Southern blots is described which is based on the principle outlined by Leary et al (1). This system has two main components: detection of biotinylated DNA by a two-step procedure with streptavidin and poly(alkaline phosphatase); and blocking background with Tween 20. 32fg and 80fg of lambda phage DNA was detected on dot and Southern blot hybridizations respectively. 150fg of beta-globin was detected on Southern blots of genomic DNA. This method is fast, reproducible and can detect single copy genes in 0.25 micrograms genomic DNA on Southern blots.

摘要

本文描述了一种基于Leary等人（1）概述的原理，用于在斑点印迹和Southern印迹上检测生物素化DNA探针的灵敏方法。该系统有两个主要组成部分：通过与链霉亲和素和聚（碱性磷酸酶）的两步法检测生物素化DNA；以及用吐温20阻断背景。在斑点印迹和Southern印迹杂交中分别检测到32fg和80fg的λ噬菌体DNA。在基因组DNA的Southern印迹上检测到150fg的β-珠蛋白。该方法快速、可重复，并且可以在Southern印迹上检测0.25微克基因组DNA中的单拷贝基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/97ab69df74be/nar00316-0178-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/b21de2127a05/nar00316-0175-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/651ecfd96e71/nar00316-0176-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/97ab69df74be/nar00316-0178-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/b21de2127a05/nar00316-0175-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/651ecfd96e71/nar00316-0176-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a692/322111/97ab69df74be/nar00316-0178-a.jpg

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本文引用的文献

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The use of the avidin-biotin complex as a tool in molecular biology.抗生物素蛋白-生物素复合物在分子生物学中的应用。
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