Aquaculture 2025

March 6 - 10, 2025

New Orleans, Louisiana USA

SIMPLIFIED WORKFLOW FOR FASTER GENOTYPING-BY-SEQUENCING USING PARTIAL COMBINATORIAL DUAL BARCODES AND OPTIMIZED ANALYSIS PIPELINE

Jennifer Lavoie*, Douglas Gibbons, Angela Burrell, Krishna Reddy Gujjula, Tia Dunbar, Kellsie Kice, Gail Ulloa, Julianne Crane, Rob Hnatuk, Srinivas Chadaram

 

Thermo Fisher Scientific

2130 Woodward St, Austin, TX 78744

jennifer.lavoie@thermofisher.com

 



Targeted Genotyping by Sequencing (GBS) is a robust and cost-effective method for marker-assisted breeding and selection.   Targeted GBS provides a scalable workflow, allowing for thousands of markers to be genotyped in up to thousands of samples per day.  As GBS use expands, demand increases for higher throughput, faster turnaround times, and increased multiplexing capability.

To achieve higher throughput with less material and greater ease of use, we propose a new library prep workflow using partial combinatorial dual barcodes for use with the Ion GeneStudio™ S5 System that not only increases available barcodes, but also achieves complete, normalized libraries for up to 3072 samples in less than a day.  Additionally, we propose an analysis workflow optimized for more than 6000 samples to be sequenced and genotyped in less than 24 hours using a single Ion GeneStudio™ S5 System. These optimizations enable completion of genotyping of >30,000 samples within a 5-day week.  Here we report early testing results showing high quality results achieved using this novel workflow and analysis pipeline.

For Research Use Only. Not for use in diagnostic procedures.