CRISPR Rat Activation

Get tips on using Live and Dead Cell Assay (Abcam) to perform Live / Dead assay mammalian cells - rabbit bone marrow mesenchymal stem cells

Get tips on using QuikChange Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Human - Point mutation HeLa Rab11

Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - Rabbit synovial fibroblasts

Get tips on using QuikChange Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Dog - Point mutation MDCK Rab11-FIP2

Get tips on using Wilms' Tumor 1 (WT1) Protein, Clone 6F-H2 to perform Immunohistochemistry Wilms Tumor 1 (WT1) - Rabbit Mouse -NA-

Get tips on using Corning® Transwell® polycarbonate membrane cell culture inserts to perform Cell migration / Invasion cell type - RAW 264.7

Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - Raw 264.7

Get tips on using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® to perform RNA sequencing Mouse - RAW264.7

Get tips on using Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System to perform Reporter gene assay β-galactosidase substrates - RAW 264.7

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.