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Team:MIT/MaterialsAndMethods
From 2012.igem.org
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<h2><b> Room Temperature Annealing </b><h2> | <h2><b> Room Temperature Annealing </b><h2> | ||
<OL> | <OL> | ||
| - | <LI>Preheat the heat block to | + | <LI>Preheat the heat block to 95°C, add water to the well you will be using.</LI> |
| - | <LI>Put a piece of styrofoam (covered with foil) on the heat block. Make sure the temperature is still at | + | <LI>Put a piece of styrofoam (covered with foil) on the heat block. Make sure the temperature is still at 95°C, and doesn't jump</LI> |
<LI>Mix gate and output in a 1.2:1 ratio in a tube, put it in the heat block.</LI> | <LI>Mix gate and output in a 1.2:1 ratio in a tube, put it in the heat block.</LI> | ||
<LI>Cover again with the styrofoam, set the heat block to RT.</LI> | <LI>Cover again with the styrofoam, set the heat block to RT.</LI> | ||
| - | <LI>5. Wait until the block cools. It will probably not reach RT, but will be at around | + | <LI>5. Wait until the block cools. It will probably not reach RT, but will be at around 30°C after a few hours. Turn the block off completely and wait for a bit (1h).</LI> |
</OL> | </OL> | ||
| + | | ||
| + | N.B. It may be possible to anneal using a lower maximum temperature (say, 80°C), as long as it is higher than the melting temperature of the oligos. | ||
| | ||
*Adapted from Qian L, Winfree E. <i> Scaling up digital circuit computation with DNA displacement cascades. </i>, Science. 2011 Jun 3;332(6034):1196-201. | *Adapted from Qian L, Winfree E. <i> Scaling up digital circuit computation with DNA displacement cascades. </i>, Science. 2011 Jun 3;332(6034):1196-201. | ||
Revision as of 14:16, 10 September 2012
Gate Anneals
Room Temperature Annealing
- Mix gate and output in a 1.2:1 ratio in a tube.
- Cover the tube in foil, if you didn't use an opaque/black tube.
- Leave the tube on a bench top overnight.
Room Temperature Annealing
- Preheat the heat block to 95°C, add water to the well you will be using.
- Put a piece of styrofoam (covered with foil) on the heat block. Make sure the temperature is still at 95°C, and doesn't jump
- Mix gate and output in a 1.2:1 ratio in a tube, put it in the heat block.
- Cover again with the styrofoam, set the heat block to RT.
- 5. Wait until the block cools. It will probably not reach RT, but will be at around 30°C after a few hours. Turn the block off completely and wait for a bit (1h).
N.B. It may be possible to anneal using a lower maximum temperature (say, 80°C), as long as it is higher than the melting temperature of the oligos.
*Adapted from Qian L, Winfree E. Scaling up digital circuit computation with DNA displacement cascades. , Science. 2011 Jun 3;332(6034):1196-201.
- Mix gate and output in a 1.2:1 ratio in a tube.
- Cover the tube in foil, if you didn't use an opaque/black tube.
- Leave the tube on a bench top overnight.
Room Temperature Annealing
- Preheat the heat block to 95°C, add water to the well you will be using.
- Put a piece of styrofoam (covered with foil) on the heat block. Make sure the temperature is still at 95°C, and doesn't jump
- Mix gate and output in a 1.2:1 ratio in a tube, put it in the heat block.
- Cover again with the styrofoam, set the heat block to RT.
- 5. Wait until the block cools. It will probably not reach RT, but will be at around 30°C after a few hours. Turn the block off completely and wait for a bit (1h).
N.B. It may be possible to anneal using a lower maximum temperature (say, 80°C), as long as it is higher than the melting temperature of the oligos.
*Adapted from Qian L, Winfree E. Scaling up digital circuit computation with DNA displacement cascades. , Science. 2011 Jun 3;332(6034):1196-201.
- Preheat the heat block to 95°C, add water to the well you will be using.
- Put a piece of styrofoam (covered with foil) on the heat block. Make sure the temperature is still at 95°C, and doesn't jump
- Mix gate and output in a 1.2:1 ratio in a tube, put it in the heat block.
- Cover again with the styrofoam, set the heat block to RT.
- 5. Wait until the block cools. It will probably not reach RT, but will be at around 30°C after a few hours. Turn the block off completely and wait for a bit (1h).
