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Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat microglia

Get tips on using Lipofectamine™ 3000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat astrocytes

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human osteoblasts

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human chondrocytes

Get tips on using Lipofectamine™ 3000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human chondrocytes

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 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.

Get tips on using Wizard® Plus Midipreps DNA Purification System Technical Bulletin to perform Plasmid Isolation Proteus mirabilis

Get tips on using Xfect™ Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human pulmonary artery smooth muscle cells (HPASMC)

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat aortic smooth muscle cells (rASMC)