| Reagent | Volume (μL) per reaction | Stock concentration |
|---|---|---|
| Nuclease-free water | 8.125 | – |
| Phusion HF Buffer (NEB) | 2.5 | 5x |
| 10 mM dNTPs | 0.25 | 10mM |
| Barcode Forward i5/Reverse i7 mixed Primers∗ | 1 | 5mM |
| Polymerase | 0.125 | 1U/uL |
| PCR 1 | 0.5 | 1x |
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