1) Create 100uM stock solutions of each oligo

2) Create a 20uL solution of the following:

  • 2 uL of 100uM stock solution of top oligo
  • 2 uL of 100uM stock solution of complementary bottom oligo
  • 2 uL T4 DNA ligase buffer
  • 1 uL T4 poly nuclease kinase (T4PNK)
  • 13 uL water

3) Put in thermocycler and run the following program:

  1. 37°C for 30 mins
  2. 95°C for 5 min
  3. Ramp to 25°C by 0.1% (0.1°C per second) and keep at 25°C for 5 min
  4. End at 4°C
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