Protein expression and purification Mammalian cells HEK 293

Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)

Get tips on using QIAamp DNA Mini Kit to perform DNA isolation / purification Cells - Primary cells Cyst-derived kidney epithelial cells

Get tips on using QIAamp DNA FFPE Tissue Kit to perform DNA isolation / purification Cells - Primary cells Pseudomyxoma peritonei (PMP) cells

Acid phosphatase detection heavily relies on determining the concentration of tartrate-resistant acid phosphatase (TRAP) in the sample. Hence, sample preparation is very crucial and it should be done strictly as per kit manufacturer instructions to avoid any inconsistency and poor sensitivity.

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