Site Directed Mutagenesis (SDM) Human Point mutation PC-3

Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human prenatal cardiac progenator cells

Get tips on using TUNEL Assay Kit - BrdU-Red to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using TransIT®-LT1 Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - Cal 27 cells Polymer / lipid

Get tips on using FlowCellect Autophagy LC3 antibody based kit to perform Autophagy assay cell type - MT-2 (human T cell leukaemia)

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - MT-2 (human T cell leukaemia)

Get tips on using LC3B (D11) XP® Rabbit mAb to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)

Get tips on using Zombie Aqua™ Fixable Viability Kit to perform Live / Dead assay mammalian cells - human peripheral blood mononuclear 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.