IGEM:Harvard/2006/DNA nanostructures/Protocols: Difference between revisions
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''See also [[IGEM:Harvard/2006/Protocols|iGEM Harvard 2006 protocols]]'' | |||
'''Aptamer-decorated Nanostructure Binding Assay''' | '''Aptamer-decorated Nanostructure Binding Assay''' | ||
Potential protocol for a 2 {{ul}} incubation reaction (revised with Dr. Shih's suggestions) | Potential protocol for a 2 {{ul}} incubation reaction (revised with Dr. Shih's suggestions) | ||
* In a 0.2 mL PCR tube, mix: | * In a 0.2 mL PCR tube, mix: | ||
** 0.5 {{ul}} of 4x (not 5x) [[IGEM:Harvard/2006/Stock_solutions#Bock.27s_selection_buffer|Bock's selection buffer]] | ** 0.5 {{ul}} of 4x (not 5x) [[IGEM:Harvard/2006/Stock_solutions#Bock.27s_selection_buffer|Bock's selection buffer]] | ||
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* Incubate at room temperature for 30 min. | * Incubate at room temperature for 30 min. | ||
* Load onto a non-denaturing polyacrylamide gel (10% to 20% gradient) at 4{{c}} | * Load onto a non-denaturing polyacrylamide gel (10% to 20% gradient) at 4{{c}} | ||
** Run at 130 V for approximately 90 min. | |||
** Liu runs at 25 mA for 48 h. | ** Liu runs at 25 mA for 48 h. | ||
Latest revision as of 09:12, 2 August 2006
See also iGEM Harvard 2006 protocols
Aptamer-decorated Nanostructure Binding Assay Potential protocol for a 2 μL incubation reaction (revised with Dr. Shih's suggestions)
- In a 0.2 mL PCR tube, mix:
- 0.5 μL of 4x (not 5x) Bock's selection buffer
- 1.0 μL of 2 μM aptamers (final concentration: 1.0 μM = 2 pmol)
- 0.5 μL of 2 μM thrombin (final concentration: 0.5 μM = 1 pmol)
- OR in a 0.2 mL PCR tube, mix:
- 0.5 μL of 4x (not 5x) Bock's selection buffer
- 0.5 μL of 2 μM aptamers (final concentration: 0.5 μM = 1 pmol)
- 1.0 μL of 2 μM thrombin (final concentration: 1.0 μM = 2 pmol)
- Alternative mix: Liu uses 10 pmol of DNA (1 μL of 10 μM) and varies thrombin amount from 2 pmol (1 μL of 0.2x thrombin working stock) to 100 pmol (1 μL 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[[:Category:{{{1}}}|{{{1}}}]]
- Run at 130 V for approximately 90 min.
- Liu runs at 25 mA for 48 h.
