| Adapter-ligated DNA fragments | 23 μL |
| NEBNext High Fidelity 2× PCR Master Mix (blue) | 25 μL |
| Index primer* (blue) | 1 μL |
| Universal PCR primer* (blue) | 1 μL |
| Total volume | 50 μL |
*The primers are provided in NEBNext Multiplex Oligos for Illumina.
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