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

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

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.

Keywords: DNA extraction; DNA isotope labeling; PCR; isotope recombination; silicon chip; surface coating.

Grants and funding

This work was partly supported by the FP7 EU Network of Excellence, Nanobeams n°500440 to Henri-Noël Migeon.