Oligo annealing

Procedure

1. Prepare a buffer. It's a good idea to use a buffer that is similar to where you will use your annealed fragment. Common options:
   • 10 mM Tris, 50 mM NaCl, 1 mM EDTA (pH 8.0)
   • 60 mM KCl, 6 mM HEPES (pH 7.5), 0.2 mM MgCl2
   • 100 mM NaPO4, 150 mM NaCl, 1 mM EDTA (pH 7.5)
   • Duplex Buffer: 100 mM K acetate, 30 mM HEPES (pH 7.5)
2. Run annealing in a thermocycler:
   1. 95C for 5 min
   2. -0.1 C/s until the sample gets to room temperature (or 4C)

Resources

• ThermoFisher
• IDT
• Metabion