Prepare for Pooling
Record the index information for each sample before starting the library prep. The preplated index adapter plates provided with these kits contain a unique combination of 10 bp dual i5 and i7 indexes per well. There are a total of 96 unique dual index combinations per plate. Each plate contains a different set of unique dual indexes. For information on the tools compatible with your sequencing system, visit the product compatibility page on the Illumina support site.
| • | For low-plexity pooling strategies (2-plex to 9-plex), refer to the Index Adapters Pooling Guide. |
| • | For index adapter sequences and information about recording the sequences, refer to Illumina Adapter Sequences. |
Supported Enrichment Plexities
Illumina DNA Prep with Exome 2.5 Enrichment reagents are configured and tested at 12-plex enrichment plexity. Although lower enrichment plexities are possible, some plexities require additional pre-enrichment library prep and enrichment probe panel reagents.
Obtaining suitable enrichment yields for nonstandard enrichment plexities might require additional optimization.
| • | Enrichment plexity—The number of pre-enrichment libraries (12 recommended, 1-11 optional) pooled together in one enrichment reaction for hybridization with the Twist BioScience for Illumina Exome 2.5 Panel and the optional Twist Bioscience for Illumina Mitochondrial Panel. For example, combining 12 pre-enrichment libraries together creates a 12-plex enrichment pool. |
| • | Enrichment reaction —The number of unique enrichment reaction preparations, regardless of the number of pre-enrichment libraries pooled per reaction (8 reactions provided in these kits). |
To calculate the total number of post-enrichment libraries, multiply the enrichment plexity per reaction by the number of enrichment reactions. For example, a single enrichment reaction of a 12-plex enrichment pool produces a pool of 12 post-enrichment libraries.
When pooling pre-enrichment libraries, Illumina DNA Prep with Exome 2.5 Enrichment reagents support the number of enrichment reactions and plexity indicated.
|
Illumina DNA Prep with Exome 2.5 Enrichment Reagents |
Enrichment Reactions |
Enrichment Plexity |
|---|---|---|
|
96-sample kit |
8 reactions |
12-plex |
|
Twist Bioscience for Illumina Mitochondrial Panel (32 µl oligo panel)² |
8 reactions¹ |
12-plex |
|
Illumina Custom Enrichment Panel v2 (32 μl, 120 bp)² |
8 reactions |
12-plex |
¹ Mitochondrial Panel may be diluted before use. Read the protocol carefully to determine the volume and dilution of oligo panel needed for your experiment.
² Sold separately. Refer to Product Contents for descriptions and catalog numbers.
