rna-isolation-purification-cells-primary-human-carotid-artery-endothelial-cells

Get tips on using High Pure RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized A2780

Get tips on using M-PER™ Mammalian Protein Extraction Reagent to perform Protein isolation Tissue - Human aortic endothelial cells

Get tips on using Cytoplasmic and Nuclear RNA Purification Kit to perform RNA isolation / purification Cells - immortalized Akata

Get tips on using Cytoplasmic and Nuclear RNA Purification Kit to perform RNA isolation / purification Cells - immortalized JY

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Primary cells CD8+ T cells

Get tips on using Nucleofector™ Kits for Human T Cells to perform DNA transfection Mammalian cells - Primary cells CD4+ T cells

Get tips on using Gentra Puregene Blood Kit to perform DNA isolation / purification Cells - Primary cells CD4+ T cells

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.