Get tips on using MEMα with L-Glutamine, Phenol Red, Sodium Pyruvate and Nucleosides to perform Stem cell Differentiation media mPericytes differentiation into Osteogenic cells
Get tips on using Gibco™DMEM, low glucose, pyruvate to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells
Get tips on using AmpFLSTR™ Identifiler™ Plus PCR Amplification Kit to perform Cell line authentication Human iPSC cells derived from peripheral blood mononuclear cells
Get tips on using EpiTect Methyl II PCR Assays to perform DNA methylation profiling Gene specific profiling - MCF-7 FOXA2
RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. 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 the 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.
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™KnockOut™ DMEM to perform Stem cell Differentiation media hiPSCs or hESCs differentiation to Embryoid body (EB)
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 Gibco™KnockOut™ DMEM/F-12 to perform Stem cell Differentiation media hiPSCs or hPSCs differentiation into trophoblasts
Get tips on using STEMdiff™ APEL™ 2-LI Medium to perform Stem cell Differentiation media hESCs or hiPSCs differentiation into Cardiomyocytes
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