RNA sequencing

Get tips on using TriPure Isolation Reagent to perform RNA isolation / purification Tissue - Mouse Muscle

Get tips on using TriPure Isolation Reagent to perform RNA isolation / purification Tissue - Mouse Liver

Get tips on using TriPure Isolation Reagent to perform RNA isolation / purification Cells - immortalized C6

Get tips on using QIAzol Lysis Reagent to perform RNA isolation / purification Cells - immortalized CT26

Get tips on using TriPure Isolation Reagent to perform RNA isolation / purification Cells - immortalized C2C12

Get tips on using TriPure Isolation Reagent to perform RNA isolation / purification Cells - immortalized U937

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.

Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Cells - primary mouse dorsal root ganglion neurons

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary lamb pulmonary artery endothelial cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human placental venous endothelial cells