rna-isolation-purification-tissue-mouse-lung

Get tips on using CD31 (PECAM-1) Monoclonal Antibody (390), Biotin, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD31/Pecam-1

Get tips on using anti-p62 / SQSTM1 (C-terminus) guinea pig polyclonal, serum to perform Autophagy assay cell type - MEFs (mouse embryonic fibroblasts)

Get tips on using LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit to perform Live / Dead assay mammalian cells - mouse, T-cell

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using CD184 (CXCR4) Monoclonal Antibody (2B11), Alexa Fluor 488, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD184/CXCR4

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Rat_Circumvallate papillae

Get tips on using TRI Reagent™ Solution to perform Protein isolation Bacteria - Vibrio cholerae

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.