Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM)/ F12 1:1: Liquid to perform 3D Cell Culture Media U87MG cells- glioblastoma spheres
Get tips on using HyClone Dulbecco's Modified Eagle Medium (DMEM) with low glucose: Liquid to perform Stem cell culture media hAdipose derived stem cells
Get tips on using QIA33 | FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme to perform Apoptosis assay cell type - Human endometrial stromal 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.
Get tips on using Lipofectamine® RNAiMAX Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - Jurkat cells Lipofectamine
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 Gibco™DMEM/F-12, no glutamine to perform Stem cell Differentiation media hPSCs or iPSCs differentiation into Lung progenitor cells
Get tips on using CytoSelect™ 24-Well Wound Healing Assay to perform Wound healing assay cell type - human gHMVEC (glioma human microvascular endothelial 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 AmpFLSTR™ Identifiler™ Direct PCR Amplification Kit to perform Cell line authentication Human iPSC cells derived from human dermal fibroblasts
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