Supplementary Figure S1. HPV capture methodology from Targeted, Deep Sequencing Reveals Full Methylation Profiles of Multiple HPV Types and Potential Biomarkers for Cervical Cancer Progression
Year: 2023
Type: supplementary-materials
Abstract: (1) Genomic DNA (gDNA) was sheared into ~300 bp fragments that were end-repaired, adenylated, and ligated to Illumina TruSeq adapters. (2) The prepared library was hybridized with Agilent's Custom HPV Capture Library and IDT's xGen Lockdown Probes. (3) Streptavidin-coated beads were used to target and separate the 5'-biotinylated end of the bound hybrids. Unbound gDNA was washed away, and captured hybrids were eluted. (4) For methylation analysis, samples were treated with BS, which converts unmethylated cytosines to uracil. (5) Captured hybrids were PCR-amplified with primers specific to the Illumnia TruSeq Adapters. Finally, libraries were indexed to provide each with a unique 6 base pair code. Pooled libraries were sequenced using the Illumina HiSeq 2000 workflow.
Language: English
Subfield: Epidemiology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4361901260