IGEM:Harvard/2006/DNA nanostructures/Protocols

 Project Overview Designs Notebook Protocols Presentations Literature  

See also iGEM Harvard 2006 protocols

Aptamer-decorated Nanostructure Binding Assay Potential protocol for a 2 incubation reaction (revised with Dr. Shih's suggestions)
 * In a 0.2 mL PCR tube, mix:
 * 0.5 of 4x (not 5x) Bock's selection buffer
 * 1.0 of 2  aptamers (final concentration: 1.0  = 2 pmol)
 * 0.5 of 2  thrombin (final concentration: 0.5  = 1 pmol)
 * OR in a 0.2 mL PCR tube, mix:
 * 0.5 of 4x (not 5x) Bock's selection buffer
 * 0.5 of 2  aptamers (final concentration: 0.5  = 1 pmol)
 * 1.0 of 2  thrombin (final concentration: 1.0  = 2 pmol)
 * Alternative mix: Liu uses 10 pmol of DNA (1 of 10 ) and varies thrombin amount from 2 pmol (1  of 0.2x thrombin working stock) to 100 pmol (1  of 10x thrombin working stock)
 * Incubate at room temperature for 30 min.
 * Load onto a non-denaturing polyacrylamide gel (10% to 20% gradient) at 4
 * Run at 130 V for approximately 90 min.
 * Liu runs at 25 mA for 48 h.