High-throughput single-nucleus DNA sequencing workflow for solid pancreatic tumors

The Mission Bio Tapestri platform was utilized to overcome the limitations of bulk sequencing in solid tumors with low cellularity, allowing high-throughput targeted snDNA-seq.

Method: 38 snap-frozen tissue samples from 16 pancreatic ductal adenocarcinoma (PDAC) patients underwent an automated 30-minute nuclei extraction. The single nuclei were then encapsulated using the Tapestri microfluidic microdroplet platform for targeted multiplex PCR amplification and library preparation.

Detection & Benefits: Tapestri bypassed the “mixed signals” of bulk sequencing caused by high stromal contamination. It successfully captured an average of 2,867 single nuclei per sample with >80X coverage depth. This enabled the precise resolution of subclones, accurate tracking of phylogenetic evolution from primary tumors to metastases, and the clear detection of zygosity and co-occurrence of key driver mutations (e.g., KRAS, TP53, SMAD4) at single-cell resolution.

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