Amplify Exome-Enriched Library Pools

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This step uses PCR to amplify the enriched whole exome library pools.

• EPM (Enhanced PCR Mix)
• PPC (PCR Primer Cocktail)
• [Plate] Microseal 'B' adhesive seal
• [Tube] 8-tube strip caps
1. Prepare the following consumables:

Item

Storage

Instructions

EPM

‑25°C to ‑15°C

Thaw on ice. Invert to mix, then centrifuge briefly.

PPC

‑25°C to ‑15°C

Thaw on ice. Invert to mix, then centrifuge briefly.
2. Save the following AMP program on the thermal cycler.
• Choose the preheat lid option and set to 100°C
• Set the reaction volume to 50 μl
• 98°C for 45 seconds
• 12 cycles of:
• 98°C for 30 seconds
• 60°C for 30 seconds
• 72°C for 30 seconds
• 72°C for 5 minutes
• Hold at 10°C

Total running time is ~35 minutes.

1. Add 5 μl PPC to each well or tube.
2. Add 20 µl EPM to each well or tube and mix thoroughly as follows.
• [Plate] Seal plate and shake at 1200 rpm for 1 minute.
• [Tube] Pipette 10 times to mix, and then cap the 8‑tube strip.
3. Centrifuge the plate or 8-tube strip at 280 × g for 30 seconds.
4. Place on the preprogrammed thermal cycler and run the AMP program.

SAFE STOPPING POINT

If you are stopping, store at 2°C to 8°C for up to two days. Alternatively, leave on the thermal cycler for up to 24 hours at 10°C.