Index Adaptor and Index Primer Pooling Guidelines (NEB #E9500 and #E9530)

To download a sample sheet with the index sequences, refer to the Usage Guidelines located in the “Protocols, Manuals & Usage” tab on the product page for the NEBNext Direct Genotyping Solution Target Enrichment Kit (NEB #E9530), www.neb.com/E9530

Tables 3.1 and 3.2 list valid barcode combinations that can be sequenced together based on color balance guidelines for lllumina sequencers. Low-complexity barcode combinations that do not maintain the recommended color balance could lead to failed sequencing of the sample indexes.

Table 3.1 lists some valid indexed 5 ́ adaptor combinations (up to 8-plex) that can be sequenced together for the inline sample index. For combinations > 8, choose any column and add any additional combinations as needed.

Table 3.2 lists some valid indexed primer combinations (up to 4-plex) that can be sequenced together for the pool index in the Illumina i7 site. For combinations > 4 choose any 4-plex combination and add any other index as needed.

Table 3.1: Valid index 5 ́ adaptor combinations for NEB #E9500B-S.

Table 3.2: Valid index primer combinations for NEB #E9530B-S.