rna-isolation-purification-tissue-mouse-lung

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - mouse microglia

Get tips on using LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit to perform Live / Dead assay mammalian cells - mouse keratinocytes

Get tips on using CD137 (4-1BB) Monoclonal Antibody (17B5), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD137

Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80

Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80

Get tips on using HTRA2 MISSION shRNA Lentiviral Transduction Particles HtrA serine peptidase 2 to perform shRNA gene silencing Mouse - FL83B HtrA2

Get tips on using D-MEM (High Glucose) with L-Glutamine and Phenol Red to perform Stem cell culture media Mouse pericytes

Get tips on using MitoSOX™ Red Mitochondrial Superoxide Indicator, for live-cell imaging to perform ROS assay cell type - mouse cardiomyocytes

RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid-mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time-consuming, but provide a more permanent expression of RNAi in the cells and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines.

RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with the desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid-mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time-consuming, but provide a more permanent expression of RNAi in the cells and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines.