Pooling Preparation

When pooling libraries, record information about your samples before starting library prep. Use a recording tool compatible with your sequencing system and libraries. For compatibility information, refer to the Illumina Stranded mRNA Prep, Ligation support pages or the support pages for your system.

The protocol uses lllumina RNA UD Indexes to index libraries in two separate steps, first adding anchors and then the UD indexes. The UD index primers add distinct Index 1 (i7) and Index 2 (i5) sequences to each end of a fragment. Each index sequence is 10 bp long.

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For strategies on forming low-plex, color-balanced pools, refer to the Index Adapters Pooling Guide.

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For index adapter sequences and how to record them, refer to Illumina Adapter Sequences.