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How do I anneal complementary oligos?

  1. Dissolve the lyophilised oligonucleotide in water or TE. Use the concentrations of your oligonucleotide solutions stated on the oligonucleotide synthesis report.
      
  2. Add the following components together:

    Stock                                             Final Concentration
    Oligonucleotide 1                             100 nmole/ml
    Oligonucleotide 2                             100 nmole/ml
    10x annealing buffer*                      1x
    Nuclease-free water                          to appropriate volume

    *10x annealing buffer: 100 mM Tris-HCI, pH 7.5,1 M NaCI and 10 mM EDTA.
       
  3. Heat the oligo solution to a temperature 10°C higher than the calculated melting temperature. Maintain the temperature for 10 minutes. Remove the solution from the heating block/water bath and allow it to cool slowly to room temperature on the bench (approximately 1 hour).

  4. Store the annealed oligos at -20°C as recommended.

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