Supporting materials Universal Split Spinach Aptamer (USSA) for Nucleic Acid Analysis and DNA Computation Nanami Kikuchi 1 and Dmitry M. Kolpashchikov 1,2 1 Chemistry Department, University of Central Florida, Orlando, 32816, Florida, USA 2 National Center for Forensic Science and Burnett School of Biomedical Sciences, University of Central Florida, Orlando, 32816, Florida, USA Table of content 1) Materials and instruments 2) Table S1: oligonucleotides used in this study 3) Detailed Experimental Procedure 4) Figure S1. Design of USSA probe 5) Figure S2-S4. Suboptimal designs of USSA analyzing Mtb. F11/KZN analyte 6) Figure S5. Analysis of long analyte using USSA probe 7) References 1. Materials and instruments. All oligonucleotides were purchased from Integrated DNA Technologies, Inc. (Coralville, IA). DNAse/RNAse free water was purchased from Fisher Scientific and used for all assays including buffers, and for dissolution of oligonucleotides. Concentrations of oligonucleotide were determined based on UV light absorption at 260 nm. DFHBI was purchased from Lucerna, Inc. (New York, NY), KCl and MgCl 2 were purchased from Fisher Scientific. Trizma Hydrochloride (Tris-HCl), pH 7.40 was purchased from Sigma Aldrich. Two 2× Spinach buffer were prepared: 2  Spinach Buffer 1 (20 mM Tris-Cl, pH 7.4, 200 mM KCl, 10 mM MgCl 2 ); 2  Spinach Buffer 2 (20 mM Tris-HCl, pH 7.4, 200 mM KCl, 100 mM MgCl 2 ). All fluorescent spectra were taken using Fluorescence Spectrometer LS55 (PerkinElmer). Otherwise noted, excitation wavelength was set to 450 nm and emission was taken at 500 nm. Electronic Supplementary Material (ESI) for ChemComm. This journal is © The Royal Society of Chemistry 2017