siRNA / miRNA gene silencing Human Primary Human Aortic Endothelial Cells

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 DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma

Get tips on using Agilent DNA 1000 Kit Bioanalyzer DNA Analysis Part Number:5067-1504 to perform Cell line authentication Human lung carcinoma cell line NCI-H1299

Get tips on using Q5® Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Human - Point mutation LNCaP Androgen Receptor splice variant (AR-V)

Get tips on using Reactive Oxygen Species (ROS) Detection Assay Kit to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma

Get tips on using Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay to perform TUNEL assay cell type - A549, NCI-H460, H1299 human alveolar carcinoma