结合梳状技术和二次离子质谱法，利用（13）C和（15）N标记研究芯片上的DNA。

Combining combing and secondary ion mass spectrometry to study DNA on chips using (13)C and (15)N labeling.

作者信息

Cabin-Flaman Armelle, Monnier Anne-Francoise, Coffinier Yannick, Audinot Jean-Nicolas, Gibouin David, Wirtz Tom, Boukherroub Rabah, Migeon Henri-Noël, Bensimon Aaron, Jannière Laurent, Ripoll Camille, Norris Victor

机构信息

Equipe AMMIS, laboratoire MERCI EA 3829, faculté des Sciences et Techniques, University of Rouen, Mont-Saint-Aignan Cedex, France.

Institute of Electronics, Microelectronics and Nanotechnology (IEMN), UMR CNRS 8520, Lille1 University, Villeneuve d'Ascq, France.

出版信息

F1000Res. 2016 Jun 20;5:1437. doi: 10.12688/f1000research.8361.1. eCollection 2016.

DOI:10.12688/f1000research.8361.1

PMID:27429742

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4943295/

Abstract

Dynamic secondary ion mass spectrometry ( D-SIMS) imaging of combed DNA - the combing, imaging by SIMS or CIS method - has been developed previously using a standard NanoSIMS 50 to reveal, on the 50 nm scale, individual DNA fibers labeled with different, non-radioactive isotopes in vivo and to quantify these isotopes. This makes CIS especially suitable for determining the times, places and rates of DNA synthesis as well as the detection of the fine-scale re-arrangements of DNA and of molecules associated with combed DNA fibers. Here, we show how CIS may be extended to (13)C-labeling via the detection and quantification of the (13)C (14)N (-) recombinant ion and the use of the (13)C: (12)C ratio, we discuss how CIS might permit three successive labels, and we suggest ideas that might be explored using CIS.

摘要

梳状DNA的动态二次离子质谱（D-SIMS）成像——通过SIMS或CIS方法进行梳理和成像——此前已利用标准的纳米二次离子质谱仪50开发出来，以在50纳米尺度上揭示体内用不同非放射性同位素标记的单个DNA纤维，并对这些同位素进行定量。这使得CIS特别适用于确定DNA合成的时间、地点和速率，以及检测DNA和与梳状DNA纤维相关分子的精细重排。在这里，我们展示了如何通过检测和定量（13）C（14）N（-）重组离子以及使用（13）C:（12）C比率将CIS扩展到（13）C标记，我们讨论了CIS如何允许进行三个连续标记，并提出了可能利用CIS探索的想法。