User:David K. Barclay/Notebook/Controlling Pancreas Cell Fate Using Transcription Factors/2014/02/03

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  • Prep primers from IDT

Prep Stock Primers from IDT

  1. Write short-hand label on blue cap that makes sense
  2. Look for the nmoles on the label
  3. Do the following calculation: nmole x 10 = μL molecular biology grade H2O that you need to add to the DNA to get 100 μM
  4. Add this amount of H2O to the appropriate tube
  5. Repeat 2-4 for the remaining tubes. These will be your stock primers.
  6. Write "100 μM" on the labels of each tube.
  7. Make sure the caps are closed tightly and vortex each tube for ~30 sec. each to completely resuspend the DNA.
  8. Let the tubes sit at room temperature for 5 min. to make sure thet DNA is completely dissolved.

Make Working Solutions of Primers (20 μM)

  1. Get fresh 1.5 mL tubes. label them with the name of each stock primer. Include "20 μM" in your label.
  2. Add 80 μL molecular biology grade H2O to each tube.
  3. Add 20 μL stock primer (100 μM) to the water in each tube.
  4. Close the caps. Mix by shaking.

  1. Store all tubes (stock and working solutions) at -20°C