Pyrosequencing analyzes single-stranded DNA templates by synthesizing complementary strands. Briefly, the four nucleotides (A, T, G and C) are added sequentially by a Pyrosequencing instrument to ...

San Diego, CA, USA). First, bisulfite modification was performed with 500 ng of total DNA using an EZ DNA methylation kit (Zymo Research). Modified DNA was amplified using pyrosequencing primers ...

At least 10 separate clones were chosen for sequencing analysis. For bisulfite pyrosequencing, the PCR products were labeled with biotin and then conducted to pyrosequencing. PCR was conducted with ...

The advent of next-generation sequencing has made possible genome analysis at previously unattainable depth. Roche, Illumina and Life Technologies, among others, have developed well-established ...

Detailed protocols regarding epithelial isolation, methylation array and pyrosequencing can be found in the online supplementary materials and methods section. See online supplementary materials and ...

Cluster analysis of data was performed and corrected using Benjamini–Hochberg adjustment for multiple comparisons. Methylation was confirmed by pyrosequencing and gene expression was determined by ...