Dec 20, 2025
Version 2
Molecular Inversion Probes _User Friendly SOP V.2
- Azeza Falghoush1,
- Luis Gamboal2,
- Brandi Torrevillas1,
- Lea Starita2,
- Allison Black3,
- ksnek 1,
- Thomas Waltzek1
- 1Washington State University College of Veterinary Medicine, Pullman, WA, United States;
- 2UW Genome Sciences, University of Washington, Seattle, WA, United States;
- 3Washington State Department of Health, Shoreline, WA, United States
- PGCoE_NW_WADDL
Protocol Citation: Azeza Falghoush, Luis Gamboal, Brandi Torrevillas, Lea Starita, Allison Black, ksnek , Thomas Waltzek 2025. Molecular Inversion Probes _User Friendly SOP. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvmbb96g3p/v2Version created by Azeza Falghoush
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol at Washington Animal Disease Diagnostic Lab (WADDL) and it's working
Created: December 20, 2025
Last Modified: December 20, 2025
Protocol Integer ID: 235522
Keywords: Molecular Inversion Probes, User Friendly SOP, H5N1, viral RNA, WGS, genome of highly pathogenic avian influenza h5n1 clade, highly pathogenic avian influenza h5n1 clade, tm viral rna target capture kit, samples with high viral load, samples with lower viral load, rna virus, high viral load, sequencing library, lower viral load, genome, libraries for whole genome, whole genome, complete hpai, hpai, probe design code
Funders Acknowledgements:
Washington State Department of Health Northwest Pathogen Genomics Center of Excellence (WA DOH contract number HED29191
Grant ID: NU50CK000630
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Abstract
TM Viral RNA Target Capture Kit is used to prepare next-generation sequencing libraries for whole genome or targeted sequencing of RNA viruses. This protocol contains instructions for library preparation for sequencing on Illumina platforms. Users should refer to product labeling for the probe design code as well as the probe fill size and the design version number. The authors have tested this protocol using a probe panel designed to cover the genome of Highly Pathogenic Avian Influenza H5N1 clade 2.3.4.4b genotype B3.13. The authors have had good success recovering the complete, or nearly complete, HPAI genomes from samples with high viral loads (e.g., when Ct values are ≤30). Samples with lower viral loads are unlikely to yield the complete HPAI genome using this protocol.
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