Targeted Bisulfite Sequencing for Promoter Methylation Analysis in Severe Preterm Birth
Preterm birth (PTB), defined as the delivery of an infant before 37 weeks of gestation, results from the interaction of genetic and environmental components and constitutes a complex multifactorial syndrome. The etiology of PTB is not completely understood because of the complex interplay of genetic, environmental, and host factors.To study the methylation levels in preterm birth gene promoters, we analyzed chorioamniotic tissue from 6 severe preterm birth and 4 controls delivered at term. Genomic DNA was treated with sodium bisulfite and 12 gene promoters of preterm birth candidate genes were amplified: ACCS, BIRC3, WNT1, CXCL2, NRN1, STEAP1, EGR3, MAMDC2, CNTNAP3B, GK, PIK3AP1 and MIR155HG. Amplicons, with an average length of 1000bp, were pooled and sequenced using the MinION platform (Oxford Nanopore).