A "turn-on" and label-free fluorescent assay for the rapid detection of exonuclease III activity based on Tb(3+)-induced G-quadruplex conjugates.
A "turn-on" and label-free fluorescent assay for the specific, rapid, and sensitive detection of 3' → 5' exonuclease III activity is reported in this study. The assay is based on the Tb(3+)-promoted G-quadruplex, which lead to the enhancement of Tb(3+) fluorescence due to the energy transfer from guanines. The proposed assay is highly simple, rapid, and cost-effective, and does not require sophisticated experimental techniques such as gel-based equipment or radioactive labels. It can be used for the rapid detection of exonuclease III activity with a detection limit of 0.8 U and a RSD (n = 6) <5 %. Notably, no dye was covalently conjugated to the DNA strands, which offers the advantages of low-cost and being interference-free.
Key Lab of Analysis and Detection for Food Safety of Ministry of Education, Fujian Provincial Key Lab of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian, 350108, China.
WeiJuan
W
Yang
YaJuan
Y
Ruan
WeiHua
W
Wu
PingPing
P
Chen
LiangJun
L
Xu
FengFu
F
Fu
D001681:Biological Assay / Q000379:methods*; D002942:Circular Dichroism; D005090:Exodeoxyribonucleases / Q000032:analysis* / Q000097:blood / Q000378:metabolism*; D005453:Fluorescence; D005456:Fluorescent Dyes; D054856:G-Quadruplexes; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*; D013725:Terbium / Q000737:chemistry*; D013997:Time Factors
D005456:Fluorescent Dyes; D013725:Terbium; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III