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This study focuses on the optimization of single-stranded binding protein (SSB) concentrations and the comparative analysis of various oligonucleotide lengths (15-50 nt) and their interactions with single-stranded DNA (ssDNA). By systematically varying SSB concentrations and oligo lengths, we aim to understand the binding dynamics and efficacy of ssDNA stabilization. Results from this study have implications for DNA manipulation techniques, molecular biology procedures, and the enhancement of genetic engineering applications.
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(A)50 (T)50 (C)50 (G)50 A Oligo 3 B Oligo 1 Oligo 2 3.1 3.1 3.1 3.1 9.4 9.4 9.4 9.4 1.3 1.3 1.3 1.3 0 0 0 0 9.4 9.4 9.4 3.1 3.1 3.1 1.3 1.3 1.3 SSB (µM): SSB (µM): 0 0 0 SSB- DNA SSB-ssDNA Free DNA ssDNA 9 8 1 0 1 2 5 6 7 4 3 3 1 2 4 5 6 8 9 7 1 1 1 3 2 1 1 1 1 1 1 1 1 0 6 5 1 4 3 2 Excess cold 50-nt oligo Excess cold 20-nt oligo 15 nt 30 nt 40 nt 50 nt 20 nt C D 50 50 100 100 No SSB No SSB 3.1 3.1 3.1 3.1 3.1 9.4 9.4 9.4 9.4 9.4 10 20 10 20 1.3 1.3 1.3 1.3 1.3 SSB (µM): 0 0 0 0 0 0 0 5 SSB- DNA SSB - oligo Free DNA 50-nt oligo 20-nt oligo 5 3 1 2 4 6 7 8 9 10 1 1 1 1 1 1 1 1 1 1 2 7 8 6 3 4 5 9 2 0 5 6 8 9 10 11 12 13 3 4 7 2 1
