Primer Extension or Oligo Overlap Extension

Using Klenow Fragment (3′→5′ exo–)

This reaction will extend to single-stranded oligos with overlap to generate a complete double-stranded DNA.

The complete reaction looks like this:

First mix together oligonucleotides and water. Heat to 90°C for 2 minutes and cool at room temperature to denature and anneal the strands for extension. Add buffer and dNTPs. Mix. Add Enzyme. Mix gently. Incubate at 37°C for 10 minutes to 1 hour.

References

1. NEB website