shRNA gene silencing Mouse Prostate cancer cell lines (DU145 and PC3) CD24

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 RPMI 1640, with L-Glutamine and 25mM HEPES to perform Mammalian cell culture media VCaP

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.

Get tips on using EpiQuik Dnmt3A Assay Kit to perform DNA methylation profiling Whole genome profiling - MCF-7, MDA-MB-453 human breast cancer

Get tips on using Senescence Detection Kit - Biovision to perform Reporter gene assay β-galactosidase substrates - mouse cardiac fibroblasts

Get tips on using Senescence Detection Kit - Biovision to perform Reporter gene assay β-galactosidase substrates - mouse embryonic fibroblasts

Get tips on using SurePrint G3 Mouse Exon 4x180K Microarray Kit (165,984 Exon probes) to perform Microarray Mice - Cochlea Expression array (labelled)

Get tips on using Live/Dead Cell Double Staining Kit to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using RPMI 1640, with L-Glutamine and 25mM HEPES to perform Mammalian cell culture media THP-1

Transfection is a powerful technique that enables the study of the function of genes and gene products in cells. Based on the nature of experiments, we may need a stable DNA transfection in cells for persistent gain-of-function or loss-of-function of the target gene. For stable transfection, integration of a DNA vector into the chromosome is crucial which requires selective screening and clonal isolation. By carefully selecting a viral delivery system and related reagents we can ensure safe and highly-efficient delivery of expression constructs for high-level constitutive or inducible expression in any mammalian cell type.