Support Information: Nanoparticle Aggregation Logic Computing Controlled by DNA Branch Migration Cheng Zhang (张成),1, a) Jingjing Ma (麻晶晶),1, Schlaberg,2 Shi Liu (刘石),2 and Jin Xu (许进)1, a) b) Jing Yang (杨静),2 H. Inaki 1 Institute of Software, School of Electronics Engineering and Computer Science, Key Laboratory of High Confidence Software Technologies of Ministry of Education, Peking University, Beijing 100871, People’s Republic of China 2 North China Electric Power University, School of Control and Computer Engineering, Beijing 102206, People’s Republic of China a) b) Electronic mail: zhangcheng369@pku.edu.cn and jxu@pku.edu.cn Cheng Zhang and Jingjing Ma contributed equally to this work. 1. DNA sequences used in experiment The sequences of the oligonucleotides are listed in Table S1. NO. B abd abc 2A1 2A2 efh egh 2A4 2B1 2B2 Da Dc D1 D2 Sequence (from 5’ to 3’) TAGTGCCAGATTTTT-SH ATTCATCTAGTGCCAGAGGTATCC ATTCATCTCGCTTGTCAGGTATCC TCTGGCACTATGACAAGCGA TCTGGCACTATCGCTTGTCA CTAATCACCCTGCTTCGGGGAACTGG CTAATCACATACGCATTCCGAACTGG CCCGAAGCAGGGGGAATGCGTATG CCCTGCTTCGTCGCTTGTCA TCTGGCACTACATACGCATT GGTTTGTATTGGCATATCACTCTTTTTTTTTTTT-SH GCCTTACGAGTCTTCTTTTTTTTTTTT-SH GAGTGATATGCCAATACAAACC CACAACTCAGTA GAAGACTCGTAAGGCTACTGAGTTGTG (-SH: thoilated DNA) Table S1. DNA sequences 2. Materials The main materials and chemicals used in our experiments are as follows: gold nanoparticles (AuNPs) with diameter of 5 and 15 nm from Ted Pella; Bis(p-sulfonatophenyl)phenylphosphinedipotassium salt dihydrate (BSPP) from Strem Chemical; Tris-Borate-EDTA buffer (TBE) and Tris-Acetic-EDTA buffer (TAE) from Solarbio; disulfide protected thiolated oligonucleotides by purification of HPLC from Sangon China; agarose from BIOWEST; methylenebisacrylamide and acrylamide monomer from TCI, Stains-All from SIGMA ALORICH. 3. PAGE gel analysis of DNA strands Before starting the experiment, the hybridizations of DNA strands were tested by PAGE gel, to determine whether they would work well in strand displacement reactions. The gel experiments were performed with TAE buffer by a mini gel device (Bio-rad). Then gels were stained with Stain all for 20 min. 4. Multivalent DNA/AuNPs conjugations The key method used in this work is conjugation of DNA and AuNP. 15 nm AuNP and thiol-DNA strands were used in the experiment. The specific steps were as follows: (1) The thiol-DNA strands were activated by dithio- threitol (DTT) and then extracted twice by ethyl acetate. (2) The “activated” thiol-DNA samples were added to the AuNP solution directly to (3) Then, an “Aging process” was carried out. Sodium polyphosphate buffer (pH 7.0) was added to 0.01 M. Later, NaCl was gradually added to a concentration to 0.2 M. The samples were then incubated for 40 h. (4) Unbound DNA was removed by centrifugation at 30 min. (5) The obtained DNA/AuNP conjugates were redissolved in sodium polyphosphate The main materials are as follows: Disodium hydrogen phosphate, sodium dihydrogen phosphate, NaCl, DTT, ethyl acetate (All chemicals were of analytical grade); 15 nm AuNP from Tedpella; All DNA samples were from Shang- hai Sangon. The equipment: Alpha Imager, Anke TGL-16G centrifuge, Bio photometer (eppendorf), ELGA Ultra-Pure Water System, Sartorius electronic balance. Tne AuNPs concentration was detected using extinction coefficients: ε520 (5 nm) = 9.3 x 106 M-1cm-1. 5. Triggering the aggregation of nanoparticles In each reaction system, the volume of AuNP solution labeled by specific DNA strands was 100 μL. NaCl was added to 0.2 M, and products of DNA displacement were gradually added to the solution of AuNPs (adding small volumes for 4-8 times). At the end of the reaction, the experimental results were detected by absorbance at 520 nm. All reactions were performed in triplicate with independent samples. 6. TEM images TEM images were obtained using a Tecnai G20. To increase surface hydrophilicity, TEM grids were ionized for 20-30 s using oxygen plasma cleaner (Harrick Plasma PDC-32). Then 2-4 μL of purified sample was spotted on the surface and left on the grid for 3 min. Later, added 10 μL of 0.5X TBE buffer for washing and used filter paper to wick off excess buffer. After repeating washing for 1-2 times, grids were air-dried for analyzing. In the transmission electron micrographs, there were many typical products as shown in Fig S1. Scale bars 100 nm, 50 nm and 20 nm. In Fig S1A, simple unit: 5 nm nanoparticles surrounded one 15 nm particle to form a particle shell. In Fig S1B, many simple units aggregated to form a large cluster. Figure S1. TEM Analysis of AuNPs /DNA conjugates products