Team:MIT/MaterialsAndMethods

From 2012.igem.org

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&nbsp;<b>N.B. </b> It may be possible to anneal using a lower maximum temperature (say, 80&degC), as long as it is higher than the melting temperature of the oligos.
&nbsp;<b>N.B. </b> It may be possible to anneal using a lower maximum temperature (say, 80&degC), as long as it is higher than the melting temperature of the oligos.
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&nbsp;*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.
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Revision as of 14:48, 10 September 2012

iGEM 2012

Bacterial

  • Placeholder1
  • Placeholder2

In Vitro

  • Gate Anneals
  • Plate Reader Studies

Mammalian

  • DNA Transfection
  • RNA Transfection

Gate Anneals

Room Temperature Annealing

  1. Mix gate and output in a 1.2:1 ratio in a tube.
  2. Cover the tube in foil, if you didn't use an opaque/black tube.
  3. Leave the tube on a bench top overnight.


Room Temperature Annealing

  1. Preheat the heat block to 95&degC, add water to the well you will be using.
  2. Put a piece of styrofoam (covered with foil) on the heat block. Make sure the temperature is still at 95&degC, and doesn't jump
  3. Mix gate and output in a 1.2:1 ratio in a tube, put it in the heat block.
  4. Cover again with the styrofoam, set the heat block to RT.
  5. 5. Wait until the block cools. It will probably not reach RT, but will be at around 30&degC 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&degC), 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.

Plate Reader Studies

Kinetic Studies


 *Adapted from Qian L, Winfree E. Scaling up digital circuit computation with DNA displacement cascades. , Science. 2011 Jun 3;332(6034):1196-201.

RNA Transfection

PLACEHOLDER TEXT

RNA Transfection

PLACEHOLDER TEXT