siRNA / miRNA gene silencing Human PC3 (human prostate cancer cell line)

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Immortalized cell lines iPSC

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Primary human breast tumors-Mammospheres

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.

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.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using MISSION® shRNA SOX2 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans SOX2 lentiviral particles

Get tips on using MISSION® shRNA SOX6 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans SOX6 lentiviral particles

Get tips on using MISSION® shRNA ZEB2 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans ZEB2 lentiviral particles

Get tips on using MISSION® shRNA ZEB1 Lentiviral Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans ZEB1 lentiviral particles

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Whole genome profiling - OVCAR-3 human ovarian cancer