1. Thaw BP clonase on ice.

2. Mix 0.5 uL BP clonase with 1 uL appropriate pDONOR (typically ~150 ng/uL) and 1 uL attB-flanked PCR fragment (typically ~50 ng/uL).

3. Incubate at room temperature for 1-1.5 hours.

4. Ad 0.5 uL Proteinase K.

5. Incubate at 37 C 20 min.

6. Transform 1 uL into 10G competent cells.

7. Outgrow w/ SOC 1hr.

8. Plate on Kan, 30C overnight.

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