rna-isolation-purification-tissue-mouse-colon

Get tips on using truChIP Ultra-Low Chromatin Shearing Kit with Formaldehyde to perform ChIP Mouse - Osteoblasts

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

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

Get tips on using EZ-Magna ChIP™ HiSens Chromatin Immunoprecipitation Kit to perform ChIP Mouse - HSC

Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - Mouse embryonic fibroblasts

Get tips on using NucleoBond® Xtra Midi / Maxi to perform Plasmid Isolation Medicago truncatula BAC clone

Get tips on using NF-κB p65 (D14E12) XP® Rabbit mAb #8242 to perform Immunohistochemistry Mouse - NFκB / p65

Get tips on using OSTEOPONTIN (O-17) ANTI-HUMAN RABBIT IGG AFFINITY PURIFY to perform Immunohistochemistry Mouse - Spp1/OPN

Get tips on using DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit 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.