Nextera Rapid Capture Enrichment Protocol Guide (15075701 v01)

Hybridize Probes

Preparation

1 Save the NRC HYB program on the thermal cycler:

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Choose the preheat lid option and set to 100°C

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95°C for 10 minutes

} 18 cycles of 1 minute each, starting at 94°C, then decreasing 2°C per cycle

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Hold at 58°C

Pool Libraries

1 Combine 500 ng of each DNA library. Make sure that each library has a unique index.

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If the total volume is > 40 μl, concentrate the pooled sample to 40 μl.

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If the total volume is < 40 μl, increase the volume to 40 μl with RSB.

Procedure

1 Add the following items in the order listed to the NEH1 plate.

Item

DNA library sample or pool

EHB

CEX, EEX, or RCO

Volume (µl)

40

50

10

2 Shake at 1200 rpm for 1 minute.

3 Centrifuge at 280 × g for 1 minute.

4 Place on the thermal cycler and run the NRC HYB program.

5 Keep at the 58°C holding temperature for at least 90 minutes and up to 24 hours.

Nextera Rapid Capture Enrichment Protocol Guide

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