What question did this study set out to answer?

This protocol aims to prepare RNA virus sequencing libraries using Molecular Inversion Probes.

December 21, 20250 citationsOpen Access

Molecular Inversion Probes User Friendly SOP v2

AFAzeza FalghoushAl-Arab Medical University LGLuis Gamboal BTBrandi K. TorrevillasUniversity of Idaho

Key Points

This protocol aims to prepare RNA virus sequencing libraries using Molecular Inversion Probes.
Utilizes TM Viral RNA Target Capture Kit for library preparation.
Designed for sequencing RNA viruses on Illumina platforms.
Tests performed on probe panel for Highly Pathogenic Avian Influenza H5N1 genomes.
Successful recovery of complete HPAI genomes from samples with high viral loads.
Complete genomes unlikely from samples with low viral loads (Ct values >30).

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.

Ask AI

Helpful

Bookmark

View Full Paper