rna-isolation-purification-tissue-mouse-colon

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 DMEM/F-12 to perform 3D Cell Culture Media Mouse primary breast ephitelial cells-Mammospheres

Get tips on using DMEM/F-12 to perform 3D Cell Culture Media Mouse primary lung epithelial cells-organoids

Get tips on using DMEM/F-12 to perform 3D Cell Culture Media Mouse primary mammary ephitelial cells- organoids

Get tips on using EpiTect Bisulfite Kit to perform DNA methylation profiling Whole genome profiling - mouse primordial germ cells

Get tips on using Stealth siRNA(m) Atg16l2 to perform siRNA / miRNA gene silencing Mouse - Pancreatic Acinar cells Atg16l2

Get tips on using Cox-2 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - B16-F10 COX-2

Get tips on using Senescence Detection Kit - Biovision to perform Reporter gene assay β-galactosidase substrates - mouse cardiac fibroblasts

Get tips on using Senescence Detection Kit - Biovision to perform Reporter gene assay β-galactosidase substrates - mouse embryonic fibroblasts

Get tips on using SMARTpool: siGENOME Fancd2 siRNA to perform siRNA / miRNA gene silencing Mouse - B16 Melanoma cells FANCD2