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

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Eric S. Tvedte
Jane Michalski
Shaoji Cheng
Rayanna S. Patkus
Luke J. Tallon
Lisa Sadzewicz
Vincent M. Bruno
Joana C. Silva
David A. Rasko
Julie C. Dunning Hotopp
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[1] University of Maryland School of Medicine,Institute for Genome Sciences
[2] University of Maryland School of Medicine,Department of Microbiology and Immunology
[3] University of Pittsburgh,Department of Medicine
[4] University of Maryland School of Medicine,Greenebaum Cancer Center
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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.
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