rna-isolation-purification-cells-primary-rat-cortical-neurons

Get tips on using Iscove’s Modified Dulbecco’s Medium to perform Stem cell Differentiation media iPSCs or hESCs differentiation into cerebellar neuroepithelium (NE)

Get tips on using OxiSelect™ Intracellular ROS Assay Kit (Green Fluorescence) to perform ROS assay cell type - SH-SY5Y human neuroblastoma

Get tips on using Gibco™ DMEM/F-12, GlutaMAX™ supplement to perform Stem cell Differentiation media iPSCs or hESCs differentiation into cerebellar neuroepithelium (NE)

Get tips on using Cas9m4-VP64 to perform CRISPR Human - Activation NEUROD1

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 GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Mouse - Neuroblastoma 2a Epac1

Get tips on using CelLytic™ B Cell Lysis Reagent to perform Protein isolation Bacteria - Bacillus cellulosilyticus

Get tips on using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Mouse - Neuroblastoma 2a dibasic site, R18Q

Get tips on using pOPINE to perform Protein expression and purification Bacteria - Escherichia coli medin

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.