Overhang or Blunt End see Note

1. Prepare dephosphorylated DNA solution (at 0.1 pmol/pL; see Note 11) in 10 mM Tris-HCl, pH 9.5,1 mM spermidine, and 0.1 mMEDTA (IX buffer A).

2. Denature the DNA by placing tube in a boiling water bath for 3 min.

3. Briefly spin tube in an Eppendorf centrifuge (hill speed, 2 s), then quickly chill on ice, and rapidly add 2 pL of buffer B.

4. Add 2-5 U of polynucleotide kinase.

5. Add 3 pL of y-PPl-ATP, and sterile distilled water to a 20-pL final vol.

7. Proceed to the hybridization step (Section 3.4.; see Note 12).

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