DNA Damage Assay Human bronchial epithelial cells (hBE)

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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.

RNA RNA isolation / purification Cells immortalized OSC-C2

Get tips on using pcDNA3.1-MRP4-his6 to perform Protein Expression Eukaryotic cells - HEK293 human MRP4-his6

Products Alice J. Rothnie, Life & Health Sciences, Aston University pcDNA3.1-MRP4-his6

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - BHK-21

Products Thermo Fisher Scientific LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - mouse iPSC

Products Thermo Fisher Scientific LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - mouse splenocytes

Products Thermo Fisher Scientific LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation

Get tips on using Gibco™ DMEM, high glucose to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Products Fisher Scientific Gibco™ DMEM, high glucose

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - mouse pancreatic stellate cells

Products Cell Signaling Technology Senescence β-Galactosidase Staining Kit - Cell Signaling

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem cells

Products Cell Signaling Technology Senescence β-Galactosidase Staining Kit - Cell Signaling

Get tips on using PE Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - T-cells Mouse (CD4+ and CD8+)

Products BD Biosciences PE Annexin V Apoptosis Detection Kit I

Get tips on using ApopTag® Fluorescein In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - Islets of langerhans (Beta cells)

Products Millipore ApopTag® Fluorescein In Situ Apoptosis Detection Kit

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