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Star Republic: Guide for Biologists

Annealing of oligonucleotides

1. Mix equal mole of both complementary oligos in a 1.5 ml eppendorf tube:

oligo 1 (0.1-1 mg) 2 ml
oligo 2 (0.1-1 mg) 2 ml
10X Annealing Buffer (100 mM Tris-HCl pH8.0, 500 mM NaCl, 10 mM EDTA) 2 ml
ddH2O 14 ml

2. Place tube in a heatblock filled with water at 90 C for 5 minutes.

3. Take the heatblock out or turn the power off, allow it to cool slowly to room temperature.

4. Place on ice or 4 C until ready to use.