2.2 Sequencing of CSFR single nucleotide polymorphisms (SNPs)
Sequencing was performed using the Ion Torrent platform. CSF3R gene exon
region and exon-intro junction region primers were designed by Ampliseq
design. The
Ion
Ampliseq Library Kit, 2.0(Thermo Fisher) was used for library
construction. The Ion PGM Hi-Q OT2 Kit and Hi-Q Sequencing 200 Kit
reagents were used to perform sequencing. Then, sequencing BAM files and
variant caller files were acquired. We then performed data screening and
judged the data quality based on mutation frequency, sequencing quality
value, forward and reverse sequencing depth and chain bias. Then, SNP
sites were screened by population frequency, mutation type and mutation
function.