rna-isolation-purification-cells-immortalized-saos-2

Get tips on using TdT In Situ Apoptosis Detection Kit - Fluorescein to perform TUNEL assay cell type - Rat fibroblast-like synoviocytes

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - H9c2 rat cardiomyocytes

Get tips on using OxiSelect™ Intracellular ROS Assay Kit (Green Fluorescence) to perform ROS assay cell type - H9c2 rat cardiomyocytes

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.

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.