crispr-human-repression-akt2

Get tips on using Gibco™ MEM α, GlutaMAX™ Supplement, no nucleosides to perform Stem cell Differentiation media Human oogonial stem cells differentiation into oocytes

Get tips on using β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer to perform Reporter gene assay β-galactosidase substrates - human MSCs (mesenchymal stem cells)

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - MCF-7 human breast cancer cells

Get tips on using Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells to perform Live / Dead assay mammalian cells - FE002-SK2 human skin progenitor cells

Get tips on using FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

Get tips on using DMEM/Ham's F-12 liquid medium w/o L-Glutamine to perform Stem cell culture media Human Tendon Stem/Pluripotence cells (TSPCs)

Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

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.

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 Reactive Oxygen Species (ROS) Detection Assay Kit to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma