shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y) Connexin 43

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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.

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Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human osteoblasts

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Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary human keratinocytes

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human keratinocytes