Supplementary Information. Silver Nanoclusters Beacon as Stimuli-Responsive Versatile. Platform for Multiplex DNAs Detection and

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1 Supplementary Information Silver Nanoclusters Beacon as Stimuli-Responsive Versatile Platform for Multiplex DNAs Detection and Aptamer-substrate Complexes Sensing Guoliang Liu,,, Jingjing Li,, Da-Qian Feng, Jun-Jie Zhu,*, Wei Wang*, State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing , China. School of Chemistry and Chemical Engineering, Yancheng Institute of Technology, Yancheng , China. School of Medical Imaging, Xuzhou Medical University, Xuzhou, China. These authors contributed equally to this work. *To whom correspondence should be addressed. Fax/Tel: , S-1

2 1, 2, 3 1, 2, 3 II denaturation 6, 7, 8 annealing 6, 7, 8 I Q III 9, 10, 11 9, 10, 11 Scheme S1. Schematic illustration of the formation of molecular beacon ssdna templates which were selected as the templates for synthesizing fluorescence silver nanoclusters. S-2

3 1, 2, 3 II 6, 7, 8 I III + Ag NO 3 Na BH 4 9, 10, 11 Scheme S2. The strategy of the synthesis of fluorescence silver nanoclusters using ssdna or SNCB. S-3

4 a b 10 nm 10 nm Figure S1. HR-TEM images of 1 ssdna-ag NCs (a) or 6 silver nanoclusters beacon (SNCB) (b). S-4

5 3000 Intensity (a.u.) Figure S2. Fluorescence emission spectra of synthesized 7 SNCB probe at 12 hour (the black curve) and 6 months (the red curve). S-5

6 Intensity (a.u.) Figure S3. Fluorescence spectra of 9 ASNCB probe in absence and presence of target DNA 4. Conditions: 1 (the black curve), 9 ASNCB probe; 2 (the red curve), nm DNA 4. S-6

7 aabsorbance 1.2 b Absorbance Figure S4. (a) The UV-vis spectra of pre-prepared 9 ASNCB probe in the absence and presence of vary concentrations of target DNA 4. Conditions: 1 (The black curve), 9 ASNCB; 2 (the red curve), µm DNA 4; 3 (the blue curve), µm DNA 4. (b) The UV-vis spectra of pre-prepared 10 ASNCB probe in the absence and presence of increasing concentrations of target DNA 5. Conditions: The black curve. Conditions: 1 (The black curve), 10 ASNCB; 2 (the red curve), µm DNA 5; 3 (the blue curve), µm DNA 5. S-7

8 a Intensity (a.u.) b Intensity (a.u.) Figure S5. (a) The fluorescence excitation (black curve) and emission spectra (red curve) of 12 SNCB. (b) The fluorescence spectra of 14 ASNCB. S-8

9 1.0 Normalize (F-F 0 ) Time (min) Figure S6. Time-dependent fluorescence changes upon detecting 1 mm ATP by activatable 14 ASNCB probe (1, black curve) or activatable 17 ASNCB probe (2, red curve). S-9

10 a 2500 b Intensity (a.u.) Intensity (a.u.) Figure S7. (a) The fluorescence excitation (black curve) and emission spectra (red curve) of 13 SNCB. (b) The fluorescence spectra of 15 ASNCB. S-10

11 Table S1 Summary of the reported fluorescence DNA sensing approaches. Method Detection Detection time References limit (nm) interval (h) DNA-templated Ag NCs GO/nucleic-acid-stabilized Ag NCs GO/molecular beacons Polymerization/nicking DNA machine Releasing acridine orange from GO NIR optical tweezers Cobalt oxyhydroxide nanoflake Activatable SNCB Present study S-11

12 Table S2 Summary of the reported optical ATP sensing approaches. Method Detection Detection time References limit (µm) interval (h) Fluorescence, GO/nucleic-acid-stabilized Ag NCs Fluorescence, Aptasensor using SYBR Green I Fluorescence, Aptamer molecular beacon (MB) Colorimetric, G-quadruplex DNAzyme Fluorescence, quenching of released dye from dsdna by GO Fluorescence, activatable SNCB Present study S-12

13 Table S3 Summary of the reported optical Thrombin sensing approaches. Method Detection Detection time References limit (nm) interval (h) Fluorescence, GO/nucleic-acid-stabilized Ag NCs Fluorescence, Aptasensor using SYBR Green I Fluorescence, Aptamer biosensor Colorimetric, G-quadruplex DNAzyme Fluorescence, polymerization/nicking DNA machine Fluorescence, activatable SNCB Present study S-13

14 Table S4 Application of the proposed method for the determination of ATP in urine samples (n = 3) by 14 ASNCB probe. The urine sample was diluted for 100 times from the original spiker urine sample. The developed 14 ASNCB approach in this paper HPLC method Sample Amount Amount Amount found a R.S.D. Recovery Amount found a R.S.D. Recovery No. added added (µm) (%) (%) (µm) (%) (%) (µm) (µm) ± ± ± ± ± ± a Average value of three determinations ± standard deviation. S-14

15 Table S5 Application of the proposed method for the determination of thrombin in human serum samples (n = 3) by 15 ASNCB probe. The serum sample was diluted for 200 times. The developed 15 ASNCB approach in this paper HPLC method Sample Amount Amount Amount found a R.S.D. Recovery Amount found a R.S.D. Recovery No. added added (nm) (%) (%) (nm) (%) (%) (nm) (nm) ± ± ± ± ± ± a Average value of three determinations ± standard deviation. S-15

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