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

Get tips on using CD140a (PDGFRA) Monoclonal Antibody (APA5), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD140/PDGFR-α

Get tips on using Anti-Collagen Type VII Antibody, clone 32,-VII to perform Immunohistochemistry Collagen VII [II-32] - Mouse Human -NA-

Get tips on using CD326 (EpCAM) Monoclonal Antibody (G8.8), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD326/EpCAM

Get tips on using CD137 (4-1BB) Monoclonal Antibody (17B5), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD137

Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80

Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80

Get tips on using HTRA2 MISSION shRNA Lentiviral Transduction Particles HtrA serine peptidase 2 to perform shRNA gene silencing Mouse - FL83B HtrA2

Get tips on using miRCURY Exosome Cell/Urine/CSF Kit to perform Purification of extracellular vesicles Exosomes - Seminal plasma

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.