IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-20: Difference between revisions
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*Goal: to get cleaner purification of oligos away from nanostructures and to increase the volume of purified nanostructures we have for protection assays. | *Goal: to get cleaner purification of oligos away from nanostructures and to increase the volume of purified nanostructures we have for protection assays. | ||
* Nanostructures: | * Nanostructures: | ||
**using 30mM {{mgcl2}}, 1x oligos ( | **using 30mM {{mgcl2}}, 1x oligos (seems to work best given [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-15#3._PEG|previous results]]). | ||
**designs: c5.0.A, c5.0.C, c5.0.D [[Table_of_c5.0_Working_Stocks|key]] | **designs: c5.0.A, c5.0.C, c5.0.D [[Table_of_c5.0_Working_Stocks|key]] | ||
* PEG: 8%, 10%. Total volume in each is 100 {{ul}} | * PEG: 8%, 10%. Total volume in each is 100 {{ul}} | ||
Revision as of 09:13, 20 August 2006
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 (seems 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
