Protein expression and purification Insect cells S2

Get tips on using RIPA Lysis Buffer System to perform Protein isolation Mammalian cells - HOG

Get tips on using RIPA Lysis Buffer, 10X to perform Protein isolation Mammalian cells - STTG1

Get tips on using 2-D Quant Kit to perform Protein quantification Mammalian cells - SiHa

Get tips on using Gentra Puregene Cell Kit to perform DNA isolation / purification Cells - Primary cells HUVEC

RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. 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.

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human airway epithelial cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary mouse tracheal epithelial cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary rat marrow stromal cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human tracheal epithelial cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human skeletal muscle cells