Ligate Adapters

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• Library Adapter Mix
• Library Prep Mix A
• Nuclease-free water (kit provided)
1. Prepare the following consumables:
• Library Prep Mix A—Pipette slowly to mix, and then centrifuge briefly. Keep on ice.
2. Save the following Ligation program on the thermal cycler:
• 20°C for 20 minutes
• Ensure that the heated lid is off.
1. Combine the following volumes to prepare diluted Library Adapter Mix. For multiple samples, multiply each volume by the number of samples plus 10% overage.
• Library Adapter Mix (0.75 µl)
• Nuclease-free water (4.25 µl)

These volumes produce 5 µl per sample.

Store dilutions of the Library Adapter Mix for up to one week at -25°C to -15°C.

2. Add 5 μl diluted Library Adapter Mix to each reaction.
3. Vortex to mix, and then centrifuge briefly.
4. Add 20 μl Library Prep Mix A to each reaction.

Library Prep Mix A is viscous, pipette slowly to ensure that proper volumes are dispensed. The total volume per reaction is 75 μl.

5. Pipette 10 times at 40 μl stroke, and then briefly centrifuge.
6. Place in the thermal cycler and run the Ligation program.