随机引物法：通过随机寡核苷酸的延伸对纯化的DNA片段进行标记。

Random Priming: Labeling of Purified DNA Fragments by Extension of Random Oligonucleotides.

作者信息

Green Michael R, Sambrook Joseph

出版信息

Cold Spring Harb Protoc. 2019 Mar 1;2019(3):2019/3/pdb.prot100586. doi: 10.1101/pdb.prot100586.

Abstract

In this protocol, oligonucleotides that are heterogeneous in sequence serve as primers for the initiation of DNA synthesis on single-stranded templates. Labeled dNTPs ([α-P]dNTPs or biotin-, DIG-, or fluorescein-labeled dUTPs) are incorporated into the new DNA by the Klenow fragment of DNA polymerase I (Pol I). The method can be adapted to radiolabel DNA in slices cut from gels cast with low-melting-temperature agarose.

摘要

在本方案中，序列异质的寡核苷酸用作在单链模板上启动DNA合成的引物。标记的脱氧核苷三磷酸（[α-P]dNTP或生物素、地高辛、荧光素标记的脱氧尿苷三磷酸）通过DNA聚合酶I（Pol I）的Klenow片段掺入新合成的DNA中。该方法可适用于对用低熔点琼脂糖浇铸的凝胶切出的切片中的DNA进行放射性标记。