适用仪器

样品实验方案

简要概述

1. 准备 DNA 样本
2. 向试管中加入试剂
3. 短暂混合并离心
4. 在 15°C 下孵育 60 分钟
5. 将反应置于冰上，然后加入停止溶液并在 65°C 下加热
6. 使用前在冰上放置 5 分钟或在 4°C 下储存
7. 纯化标记的 DNA

 注：在开始实验之前，解冻所有成分。在开始标记过程之前，将所有试剂短暂涡旋至底部。

实验步骤

表 1. 各反应每管试剂组成

可以优化生物素-dUTP（组分 A）：dTTP（组分 E）的比例以获得最佳标记条件。
可以优化孵育时间以获得更好的标记。 更长的孵育时间将有助于更多的标记，但可能会缩短最终产品的尺寸。
1.向干净的（无核酸酶）0.5 mL 微型离心管或 0.2 mL PCR 管中，按表 1 中所示的顺序添加试剂。
2.通过短暂的涡旋和短暂的离心小心混合试剂。
3.将反应在 15°C 下孵育 60 分钟。
4.孵育后，将反应置于冰上。
5.要终止反应，请添加 5 µL 停止溶液并将样品加热至 65 °C。
6.使用前在冰上放置 5 分钟或在 4°C 下储存。
7.纯化标记的 DNA。

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging.
Authors: O’Sullivan, Julia and Mersaoui, Sofiane Y and Poirier, Guy and Masson, Jean-Yves
Journal: Journal of visualized experiments : JoVE (2021)

Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed.
Authors: Malayil, Leena and Chattopadhyay, Suhana and Mongodin, Emmanuel F and Sapkota, Amy R
Journal: Environmental microbiome (2021): 13

Customized optical mapping by CRISPR-Cas9 mediated DNA labeling with multiple sgRNAs.
Authors: Abid, Heba Z and Young, Eleanor and McCaffrey, Jennifer and Raseley, Kaitlin and Varapula, Dharma and Wang, Hung-Yi and Piazza, Danielle and Mell, Joshua and Xiao, Ming
Journal: Nucleic acids research (2021): e8

Fast and Efficient Postsynthetic DNA Labeling in Cells by Means of Strain-Promoted Sydnone-Alkyne Cycloadditions.
Authors: Krell, Katja and Pfeuffer, Bastian and Rönicke, Franziska and Chinoy, Zoeisha S and Favre, Camille and Friscourt, Frédéric and Wagenknecht, Hans-Achim
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2021)

Fluorescent SAM analogues for methyltransferase based DNA labeling.
Authors: Goyvaerts, Vince and Van Snick, Sven and D’Huys, Laurens and Vitale, Raffaele and Helmer Lauer, Milena and Wang, Su and Leen, Volker and Dehaen, Wim and Hofkens, Johan
Journal: Chemical communications (Cambridge, England) (2020): 3317-3320

Metabolically-active bacteria in reclaimed water and ponds revealed using bromodeoxyuridine DNA labeling coupled with 16S rRNA and shotgun sequencing.
Authors: Malayil, Leena and Ramachandran, Padmini and Chattopadhyay, Suhana and Cagle, Robin and Hittle, Lauren and Ottesen, Andrea and Mongodin, Emmanuel F and Sapkota, Amy R
Journal: Water research (2020): 116185

Tracing Baculovirus AcMNPV Infection Using a Real-Time Method Based on ANCHORTM DNA Labeling Technology.
Authors: Hinsberger, Aurélie and Graillot, Benoît and Blachère Lopez, Christine and Juliant, Sylvie and Cerutti, Martine and King, Linda A and Possee, Robert D and Gallardo, Franck and Lopez Ferber, Miguel
Journal: Viruses (2020)

Click Chemistry-Based DNA Labeling of Cells for Barcoding Applications.
Authors: Gentile, Stefan D and Griebel, Megan E and Anderson, Erik W and Underhill, Gregory H
Journal: Bioconjugate chemistry (2018): 2846-2854

Multiplexed sgRNA Expression Allows Versatile Single Nonrepetitive DNA Labeling and Endogenous Gene Regulation.
Authors: Shao, Shipeng and Chang, Lei and Sun, Yuao and Hou, Yingping and Fan, Xiaoying and Sun, Yujie
Journal: ACS synthetic biology (2018): 176-186

Real-Time Visualization and Quantification of Human Cytomegalovirus Replication in Living Cells Using the ANCHOR DNA Labeling Technology.
Authors: Mariamé, Bernard and Kappler-Gratias, Sandrine and Kappler, Martin and Balor, Stéphanie and Gallardo, Franck and Bystricky, Kerstin
Journal: Journal of virology (2018)