AgriSeq™ ULTRA Library Kit

Key product features include:

• Designed for ultra-high-throughput genotyping of up to 3072 samples in a run
• Inhibitor tolerance for better performance with direct lysate input
• Exceptional uniformity of multiplexed target amplification generates robust, reproducible results
• Simplified workflow, including direct lysis, PCR-based barcoding, and pooling of samples for clean-up, enables high-throughput sample processing
• Convenient packaging contains library normalization reagents and volumes sufficient for automated platforms

Optimized for working with crude lysates

AgriSeq ULTRA library preparation has been optimized for targeted GBS of crude lysates prepared by direct lysis of tissue samples. The combination of inhibitor tolerant master mixes, anti-inhibitors, and additive mixes enables the AgriSeq ULTRA workflow to be effective with crude lysates with variable inputs.

Excellent data quality

AgriSeq ULTRA library preparation has been optimized for targeted GBS of panels ranging from 50–2000 amplicons. With a highly efficient and specific multiplex technology, AgriSeq amplification can generate libraries with high uniformity (91.9–99.1%), percent on-target reads (92.0–99.9%), and overall genotype call rates (89.6–99.2%) based on a set of five recently designed panels ranging in size from 130–2000 amplicons across five different species. The figure below shows an average of 97.3% uniformity for a 1856-amplicon panel across 512 samples (i.e., 97.3% of bases are covered by at least 0.2X of the average read depth). Even coverage is significant in that it reduces the amount of sequencing required to achieve consistent genotype calls, thus reducing the sequencing cost per sample. In comparison, weaker multiplexing performance is reported with the competitor technology, TruSeq, which provides only 88% average uniformity and a reported 1536 maximum multiplexing capability.

High throughput sample processing with dual combinatorial barcoding

The AgriSeq ULTRA Library workflow (see figure below) can be performed in either 96- or 384-well plates for high-throughput sample processing using as little as 10 ng of DNA input per reaction. The workflow has been optimized to help minimize sample handling and ancillary reagent usage through the processing of pooled samples. This allows the workflow to be completed with minimal pipetting steps, helping reduce the potential for error, as well as simplifying automation setup on common third-party platforms such as Tecan or Hamilton systems.

Faster data analysis: genotyping large number of samples in less than 24 hrs

Standard data analysis processing includes automated quality assessment, alignment, and genotype calling using the included Torrent Suite software and Variant Caller plug-in without the need for a bioinformatician, expensive data storage, or third-party programs.

Convenient packaging containing library normalization reagents and volumes for automation

The AgriSeq ULTRA kit is offered in a large pack size (9600 reactions) to accommodate high-throughput agrigenomics projects while minimizing the cost and waste of unnecessary packaging. The kit includes not only reagents for library preparation, but all of the components required for library normalization to help achieve consistent results between samples. With a low price per sample exclusively available for agricultural customers, AgriSeq targeted GBS is an economical way to generate high-throughput genotyping data to advance your breeding programs.