IGEM:Harvard/2006/Folding DNA nanostructures: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
ShawnDouglas (talk | contribs) mNo edit summary |
ShawnDouglas (talk | contribs) mNo edit summary |
||
Line 19: | Line 19: | ||
'''Gel Analysis''' | '''Gel Analysis''' | ||
*Run 1% agarose gel, supplemented to 10 mM {{mgcl2}} | *Run 1% agarose gel, supplemented to 10 mM {{mgcl2}} | ||
*Run at low voltage (80V) for 1 | *Also supplement 1x TBE to 10 mM {{mgcl2}} | ||
*Run at low voltage (80V) for 1 hour (longer if necessary) | |||
[[Image:SMD-gel060612a.jpg|thumb|1% agarose 10 mM {{mgcl2}}]] | [[Image:SMD-gel060612a.jpg|thumb|1% agarose 10 mM {{mgcl2}}]] |
Revision as of 12:16, 12 June 2006
- working stocks
- 44 nM scaffold (20 μL)
- 0.99 μM each oligo
- goal
- 10 nM scaffold
- 100 nM each oligo
- 20 μL reaction volume
- calculations
- scaffold = (10 nM)/(44 nM) * 20 μL = 4.5 μL
- oligos = (100 nM)/(990 nM) * 20 μL = 2 μL
- reaction mixture
- 4.5 μL p7308 scaffold (44 nM)
- 2 μL oligos (990 nM)
- 2 μL 10x folding buffer (500 mM HEPES pH 7.5, 500 mM NaCl, 100 mM MgCl2)
- 11.5 μL dH2O
- annealing protocol
- 90[[:Category:{{{1}}}|{{{1}}}]] 5' → 65[[:Category:{{{1}}}|{{{1}}}]] 20' → 55[[:Category:{{{1}}}|{{{1}}}]] 20' → 45[[:Category:{{{1}}}|{{{1}}}]] 20' → 37[[:Category:{{{1}}}|{{{1}}}]] 30'
Gel Analysis
- Run 1% agarose gel, supplemented to 10 mM MgCl2
- Also supplement 1x TBE to 10 mM MgCl2
- Run at low voltage (80V) for 1 hour (longer if necessary)
Lane | Contents | Loading Buffer |
0 | 1kb DNA ladder (10 μL) | 10x loading dye (1.1 μL) |
1 | control: +scaffold -oligos (10 μL) | 10x loading dye (1.1 μL) |
2 | control: -scaffold +oligos (10 μL) | 10x loading dye (1.1 μL) |
3 | folded nanotubes (10 μL) | 10x loading dye (1.1 μL) |