Probing DNA hybridization efficiency and single base mismatch by X-ray photoelectron spectroscopy.
We demonstrated the use of X-ray photoelectron spectroscopy (XPS) to study DNA hybridization. Target DNA labeled with hexachloro-fluorescein (HEX) was hybridized to DNA arrays with four different probes. Each probe dot of the hybridized arrays was detected with XPS. The XPS Cl2p peak areas were found to decrease with an increase in mismatched bases in DNA probes. The Cl2p core-level peak area ratio of a probe perfectly matched to one, two and three base-mismatched probes accorded well with the results of conventional fluorescent imaging, which shows that XPS is a potential tool for analyzing DNA arrays. The DNA arrays' hybridization efficiency was assessed by the molar ratio of chlorine to phosphorus in a DNA strand, which was determined from the relevant XPS Cl2p and P2p core-level peak areas after hybridization. This could provide a new method to detect DNA hybridization efficiency.
Liu
Zheng-Chun
ZC
Institute of Biomedical Engineering, School of Info-Physics and Geomatics Engineering, Central South University, Changsha, 410083, China. liuzhengchunseu@126.com
Zhang
Xin
X
He
Nong-Yue
NY
Lu
Zu-Hong
ZH
Chen
Zhen-Cheng
ZC
D020137:Base Pair Mismatch; D002713:Chlorine / Q000737:chemistry; D004583:Electrons; D005453:Fluorescence; D009693:Nucleic Acid Hybridization; D013057:Spectrum Analysis / Q000379:methods*; D014965:X-Rays
D002713:Chlorine