siRNA / miRNA gene silencing Human EM-2

Get tips on using EZ-ChIP™ to perform ChIP Human - HEK 293

Get tips on using Qubit dsDNA HS Assay Kit to perform DNA quantification Human - Colorectal aenocarenoma (SW48) - paraffin embeded

Get tips on using TTF1 Monoclonal Antibody (2F4D8) to perform Immunohistochemistry Human - TTF-1

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Human - PANC-1

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Human - MCF-7

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Human - SH-SY5Y

Get tips on using MitoSOX™ Red Mitochondrial Superoxide Indicator, for live-cell imaging to perform ROS assay cell type - A549 human adenocarcinomic human alveolar basal epithelial cells

Get tips on using β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer to perform Reporter gene assay β-galactosidase substrates - SK-Hep-1

Get tips on using Hoechst 33258 to perform DNA quantification Human - MDA-MB-231

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.