IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-20

PEG precipitation

• Goal: to get cleaner purification of oligos away from nanostructures and to increase the volume of purified nanostructures we have for protection assays.
• Nanostructures:
  • using 30mM MgCl₂, 1x oligos (seem to work best given previous results.
  • designs: c5.0.A, c5.0.C, c5.0.D key
• PEG: 8%, 10%. Total volume in each is 100 μL
  • 8 % Cocktail:

     40 μL 20% PEG
     10 μL 5M NaCl
     10 μL water
     40 μL Nanostructures (add last)

• • 10 % Cocktail:

     50 μL 20% PEG
     10 μL 5M NaCl
     40 μL Nanostructures (add last)

• incubate on ice for 15 min.
• spin at 16 k rcf at 4[[:Category:{{{1}}}|{{{1}}}]] for 10 min.
• carefully pipette off supernatant
• resuspended "pellet" in 50 μL of water
  • note: add PEG first, nanostructures last; mix using tapping after everything added. let sit for ~5 min. before putting it on ice