What guidance do you have for adding a small number of guide RNA pairs to the fixed PureTarget panel?

The PureTarget 20 gene panel is at 5 µM and includes 20 pairs of gRNAs made up of 40 individual gRNAs at 0.125 µM each.

To add on custom genes to the panel, for simplicity, we recommend the following:

1. Prepare your custom gRNAs pair per gene at final 5 µM.
2. Spike into the PureTarget gRNA mix to prepare custom gRNA mixes prior to making ‘Reaction Mix 2 (RM2)’ in step 3 of the Procedure & checklist – Generating PureTarget repeat expansion panel libraries, while ensuring the total sum molarity of the final mix is 5 µM; the volumes of gRNAs you mix should be proportional to the number of guide pairs, for example add 2 µL of a 1-plex mix to 40 µL of the 20-plex PureTarget panel to keep the sum final molarity at 5 µM.
3. Do not change any of the master mix volumes in the protocol.

Please consult with PacBio field application scientist or global service and support for more specific details on library prep