June 12 (Monday)

Folding DNA Nanostructures
1. Working Stocks 44 nM scaffold (20 microL) 0.99 microM of each oligo 2. Protocol - goal: 10 nM scaffold w/ 100 nM each oligo, total rxn vol of 20 microL - calculations: scaffold = (10 nM)/(44 nM) * 20 microL = 4.5 microL oligos = (100 nM)/(990 nM) * 20 microL = 2 microL - reaction mixture: 4.5 microL p7308 scaffold 2 microL oligos 2 microL 10x folding buffer (500 mM HEPES ph 7.5, 500 mM NaCl, 100 mM MgCl2) 11.5 microL dH2O - annealing times: 90 dC, 5' 65 dC, 20' 55 dC, 20' 45 dC, 20' 37 dC, 30' - neg. controls: 1 replacing oligos w/ water, 1 replacing scaffold w/ water

Transformation
1. Plasmids R0010: lac operon promoter E7104: T7 promoter + GFP E0241: GFP 2. Protocol - let chemically competent OneShot Top10 cells thaw on ice - added appropriate DNA to cells and tapped gently to mix - let sit on ice for 20 min - heat shocked @ 42 dC for 30"   - let cool on ice for 2 min    - added 200 microL SOC media    - shook @ 37 dC for 1 hr    - pipetted and spread onto agar plates treated w/ ?    - incubated @ 37 dC overnight agar side-up