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Conformational Change Detection of DNA with the Fluorogenic. Reagent of o-Phthalaldehyde-β-Mercaptoethanol (OPAME). Yun Fei Long,†,‡ Qie Gen Liao,† ...

Supporting information for Conformational Change Detection of DNA with the Fluorogenic Reagent of o-Phthalaldehyde- -Mercaptoethanol (OPAME) Yun Fei Long,†,‡ Qie Gen Liao,† Cheng Zhi Huang,*,† Jian Ling † and Yuan Fang Li† †

CQKL-LRTA, College of Chemistry and Chemical Engineering, Southwest

University, Chongqing 400715, China.



Institute of Chemistry and Chemical

engineering, Hunan University of Science Technology, Xiangtan 411201, China Supporting Experimental Details Stock solution of fish sperm DNA (fsDNA, Shanghai Institute of Biochemistry, Academy of Science, Shanghai, China) was prepared with Milli-Q purified water. The concentration is determined according to the absorbance values at 260.0 nm by using DNA =

6600 M cm-1. (RS1) Single stranded fish sperm DNA (ssDNA) is obtained by

incubating fish sperm DNA in boiling water bath for 10 min and cooling in ice water immediately. The images were obtained by using a portable 253 nm UV lamp light source excitating the solution dipped on the silica surface, and the images of the aqueous solutions were recorded by a Nikon coolpix-4500 digital camera. Reaction between amino compound and OPAME CHO

pH 8-9.5 + R-NH2

CHO o-phthaldialdehyde

amido compound

+

R'-SH

N-R + 2H2O

Borate buffer -mercap toethanol

fluorescent

S-R'

Scheme S1. Reaction between amino compound and OPAME. The derivative is fluorescent. 1

OH

O CHO +

CHO

R'-SH +

O

HN O

OH

HO

N

-mercap toethanol

o-phthaldialdehyde

O P OH

N

N

H2N

GMP

OH

O N

pH 8-9.5

O

N

P OH

N

(1)

+ 2H2O

S-R'

O

HN

OH

HO

N O

N

CHO R'-SH

+

CHO o-phthaldialdehyde

+

N

OH

HO

OH

H2 N

N

-mercap toethanol

O O

N

HO

N

O

OH OH

pH 8-9.5

N

Borate buffer

N

CHO

N + R'-SH + H N 2

-mercap toethanol

O

N

pH 8-9.5

Borate buffer

(2)

OH + 2H2O

P

O N

cytosine

O O

N

S-R'

o-phthaldialdehyde

OH

AMP

N

CHO

P

N

O N

S-R'

Scheme S2. The reaction mechanisms between OPAME respectively with GMP(1), AMP(2), and cytosine(3).

2

(3)

Results: 1. The fluorescent spectra of OPAME and its mixture with natural and thermally denatured DNA. 500

3 1 2 3

400 300

I

F

1

200 100 0

400

450

500

550

600

/n m

Fig. S1. Fluorescent spectra of OPAME and its mixture with natural and thermally denatured DNA. 1, OPAME (black); 2, OPAME-natural fsDNA (red); 3, OPAMEthermally denatured DNA (green). Conditions: cOPAME, 1×10-3 M; cDNA, 1.0 mg l-1. 2. Fluorescence sensing scheme based on different activities of amino-group before and after the conformational change of P1 and their respective fluorescence properties.

Fig. S2. Fluorescence sensing scheme based on different activities of amino-group before and after the conformational change of P1 and their respective fluorescence properties. This information is available free of charge via the Internet at http://pubs.acs.org.

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