rna-isolation-purification-tissue-mouse-colon

Get tips on using EZ-Magna ChIP™ A/G Chromatin Immunoprecipitation Kit to perform ChIP Mouse - Liver

Get tips on using LC3B (D11) XP® Rabbit mAb to perform Autophagy assay cell type - MEFs (mouse embryonic fibroblasts)

Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - MEFs (mouse embryonic fibroblasts)

Get tips on using Recombinant Anti-CD204/MSR1 Antibody (FITC), Rabbit Monoclonal to perform Flow cytometry Anti-bodies Mouse - CD204

Get tips on using Color-coded Prestained Protein Marker, Low Range (1.7-42 kDa) #13070 to perform Protein Ladder Prestained

Get tips on using Color-coded Prestained Protein Marker, High Range (43-315 kDa) #12949 to perform Protein Ladder Prestained

Get tips on using Color-coded Prestained Protein Marker, Broad Range (10-250 kDa) #74124 to perform Protein Ladder Prestained

Get tips on using Anti-alpha smooth muscle Actin antibody to perform Immunohistochemistry Alpha smooth muscle Actin - Rabbit Mouse -NA-

Get tips on using CellROX™ Deep Red Reagent, for oxidative stress detection to perform ROS assay cell type - mouse splenocytes

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.