siRNA / miRNA gene silencing Human UCC (urothelial cancer cell lines)

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Get tips on using siGENOME Human PDCD1LG2 (80380) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - M244 PD-L2

Get tips on using Accell Human MYB (4602) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MEG-01 MYB

Get tips on using Accell Human VEGFC (7424) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MEG-01 VEGFC

Get tips on using Accell Human MYB (4602) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - EM-2 MYB

Get tips on using Accell Human VEGFC (7424) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - EM-2 VEGFC

Get tips on using siGENOME Human PPARD (5467) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - DU145 PPAR-delta

Get tips on using siGENOME Human ATG12 (9140) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - BT-20 Atg12

Get tips on using siGENOME Human RAB5A (5868) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - BT-20 Rab5a

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.