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Get tips on using MethylFlash Methylated DNA 5-mC Quantification Kit to perform DNA methylation profiling Whole genome profiling - C2C12 mouse myoblast cells

The challenge in isolating RNA from S. aureus cells is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads is considered to be the better alternative.

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

Get tips on using H2A.X Phosphorylation Assay Kit (Flow Cytometry) to perform DNA Damage Assay Human Skin Fibroblast Cell (FSK)

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat pulmonary artery smooth muscle cell (pPASMC)

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat pulmonary artery smooth muscle cell (pPASMC)

Get tips on using Cytoplasmic and Nuclear RNA Purification Kit to perform RNA isolation / purification Cells - immortalized Akata

Get tips on using Cytoplasmic and Nuclear RNA Purification Kit to perform RNA isolation / purification Cells - immortalized JY

Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized MC-9

Get tips on using High Pure RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized A2780