| Component | Amount (μL) | Final |
|---|---|---|
| Nuclease-free water | 2.5 | |
| NEBNext Ultra II Q5 2x Master Mix with 2x SYBR Green | 5 | 1X |
| Dual Index Primer Pair (from NEBNext® 96 Unique Dual Index Primer Pairs for Illumina®), 10 μM | 0.5 | 0.5 μM (0.25 μM each primer) |
| Diluted PCR#1 product | 2 | |
| Total volume | 10 |
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