Fig. 2: Thermodynamic tuning of amplification within competitive PCRs.
From: Promiscuous molecules for smarter file operations in DNA-based data storage
a Strands with 2 HD binding sites (60 bp, orange) are screened for nonspecific amplification in a competitive reaction against 0 HD strands (200 bp, red). Two stringent conditions are individually tested (top row): (left) 250 nM primer at 60 °C and (right) 125 nM at 55 °C. The promiscuous conditions individually tested (bottom row) are (left) 250 nM primer at 45 °C and (right) 500 nM at 55 °C. Gray spheres encompass strands that are expected to be amplified, and the gel electrophoresis lanes show experimental results. b A screen of a library of sequences is conducted to find sequences to be used in scaling to full files and databases. Each forward binding sequence (letters a–f) is paired with every reverse binding sequence (numbers 1–5) in the reactions described in a. Amplification tunability is defined as the difference in the ratio of mismatch (60 bp) strands to perfect match (200 bp) strands from promiscuous to stringent conditions. Positive values represent tunability in the expected direction. Tunability using annealing temperature (black) and primer concentration (gray) are shown. Source data are provided as a Source Data file.
