Get tips on using Cytoselect™ Cell Viability and Cytotoxicity Assay to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells
Get tips on using TumorTACS™ In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells
Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells
Get tips on using in situ Cell Death Detection Kit, POD to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells
Get tips on using Gibco™ DMEM/F-12, GlutaMAX™ supplement to perform Stem cell culture media Human Fetal brain-derived neural stem cells
Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - BEAS-2B human bronchial epithelial cell line
Get tips on using OxiSelect™ Intracellular ROS Assay Kit (Green Fluorescence) to perform ROS assay cell type - BEAS-2B human bronchial epithelial cell line
Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - A549, NCI-H460, H1299 human alveolar carcinoma
Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma 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.
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