Evaluation of a high-throughput, cost-effective Illumina library preparation kit

Eric S Tvedte,Julie C Dunning Hotopp,Vincent M Bruno,Joana C Silva,Luke J Tallon,David A Rasko,Shaoji Cheng,Jane Michalski,Lisa Sadzewicz,Rayanna S Patkus

doi:10.1038/s41598-021-94911-0

Eric S Tvedte, Julie C Dunning Hotopp + Show 8 more

Open Access

https://doi.org/10.1038/s41598-021-94911-0

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Abstract

Library preparation for high-throughput sequencing applications is a critical step in producing representative, unbiased sequencing data. The iGenomX Riptide High Throughput Rapid Library Prep Kit purports to provide high-quality sequencing data with lower costs compared to other Illumina library kits. To test these claims, we compared sequence data quality of Riptide libraries to libraries constructed with KAPA Hyper and NEBNext Ultra. Across several single-source genome samples, mapping performance and de novo assembly of Riptide libraries were similar to conventional libraries prepared with the same DNA. Poor performance of some libraries resulted in low sequencing depth. In particular, degraded DNA samples may be challenging to sequence with Riptide. There was little cross-well plate contamination with the overwhelming majority of reads belong to the proper source genomes. The sequencing of metagenome samples using different Riptide primer sets resulted in variable taxonomic assignment of reads. Increased adoption of the Riptide kit will decrease library preparation costs. However, this method might not be suitable for degraded DNA.

Highlights

Library preparation for high-throughput sequencing applications is a critical step in producing representative, unbiased sequencing data
Given the potential to reduce library preparation costs for labs performing high-volume genome sequencing experiments, we sought to independently test Riptide libraries for diverse organisms that are the causative agents of multiple infectious diseases
Riptide libraries were compared to widely-used Illumina libraries (KAPA Hyper, NEBNext Ultra) generated from the same DNA sample

Summary

Introduction

Library preparation for high-throughput sequencing applications is a critical step in producing representative, unbiased sequencing data. The iGenomX Riptide High Throughput Rapid Library Prep Kit purports to provide high-quality sequencing data with lower costs compared to other Illumina library kits. To test these claims, we compared sequence data quality of Riptide libraries to libraries constructed with KAPA Hyper and NEBNext Ultra. Increased adoption of the Riptide kit will decrease library preparation costs This method might not be suitable for degraded DNA. Efforts are needed to reduce labor and reagent costs in library construction while generating high-quality whole genome sequencing data. The iGenomX Riptide High Throughput Rapid Library Prep Kit (https://igenomx.com/products/) promotes ultrafast and automatable processing of biological samples to yield high-quality Illumina sequence data.

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