rna-isolation-purification-cells-primary-canine-peripheral-blood-mononuclear-cells

Get tips on using Gibco™ DMEM, high glucose, GlutaMAX™ Supplement to perform Stem cell culture media Mouse muscle satellite cells

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - K562 human leukemia cells

Get tips on using OxiSelect™ Intracellular ROS Assay Kit (Green Fluorescence) to perform ROS assay cell type - K562 human leukemia cells

Get tips on using Purified Mouse Anti-Nucleoporin p62 Clone 53/Nucleoporin p62 to perform Autophagy assay cell type - N27 dopaminergic cells

Get tips on using PE Annexin V Apoptosis Detection Kit with 7-AAD to perform Apoptosis assay cell type - Human T-cells

Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

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. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

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. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

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

Get tips on using β-Gal Reporter Gene Assay, chemiluminescent to perform Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem cells