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

Get tips on using CD11b Monoclonal Antibody (M1/70), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD11b

Get tips on using CD45 Monoclonal Antibody (30-F11), PE-Cyanine7, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD45

Get tips on using GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit to perform CRISPR Mouse - Deletion NIH 3T3 G3BP

Get tips on using GeneArt™ CRISPR Nuclease Vector with CD4 Enrichment Kit to perform CRISPR Mouse - Deletion NIH 3T3 FXR

Get tips on using Purified anti-Tubulin β-3 (TUBB3) Antibody (Previously Covance catalog# PRB-435P) to perform Immunohistochemistry Mouse - TUBB3

Get tips on using IMPACT™ Kit to perform Protein expression and purification Bacteria - Escherichia coli GmrSD

Get tips on using pGS-21a Vector to perform Protein expression and purification Bacteria - Escherichia coli p19

Get tips on using QIAexpress Type IV Kit to perform Protein tag Purification of His-tagged proteins

Get tips on using GeneRead Size Selection Kit (50) to perform Whole Genome Amplification NGS library purification

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.