Epigenome analysis of DU145 samples

Bisulphite converted DNA from DU145 cell samples were hybridized to the Illumina Infinium 850k Human Methylation Beadchip. The Illumina Infinium 850k Human DNA methylation Beadchip was used to obtain DNA methylation profiles in DU145 cell samples.Genomic DNA was isolated from DU145 cells and bisulfite conversion was performed. Add 0.1 N NaOH to the sample to denature the DNA to single-stranded DNA. After neutralization, add the whole genome amplification reagent, and incubate overnight at 37 °C. The DNA fragments were precipitated by adding isopropanol, and the fragmented DNA was enriched by centrifugation at 4 °C and purified. After the precipitated DNA is dried in the air, a hybridization buffer reagent is added to re-dissolve the DNA precipitate. Hybridize the resuspended DNA sample with the prepared chip and place it in a hybridization oven overnight. Wash away unhybridized and non-specifically hybridized DNA for subsequent staining and extension. Using the captured DNA as a template, a single-base extension reaction is performed on the chip, and a detectable label group is added to the chip to distinguish the methylation of the sample. Put the reaction-completed chip into XC4 reagent and then put it in a vacuum environment to dry for 1 hour. Put the processed chip into a scanner, use laser to excite the fluorescent group of the single-base extension product on the chip, and the scanner obtains the fluorescence emitted by the fluorescent group and generates a high-resolution picture. The data thus obtained is directly imported into GenomeStudio software for analysis, so as to obtain the methylation data of each sample. Samples included normal , PLK1 knockdown and PLK1 knockdown with over-expression UHRF1 DU145 cell samples, Each sample is repeated three times. [contributor] Oebiotech