DNA methylation profiling Gene specific profiling Rat whole pituitary glands

Get tips on using m6a rna methylation quantification kit to perform RNA quantification Coloremetric

Get tips on using EpiTect MethyLight PCR +ROX Vial Kit (200) to perform PCR Methylation specific PCR - Mammalian DNA

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Tissue - Rat Adrenal glands

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.

Get tips on using Dynabeads™ mRNA Purification Kit to perform RNA isolation / purification Tissue - Rat Adrenal glands