rna-isolation-purification-cells-immortalized-saos-2

Get tips on using 14-3-3ζ siRNA(h) to perform siRNA / miRNA gene silencing Human - Caco-2 14‐3‐3ζ

Get tips on using β-Gal Reporter Gene Assay, chemiluminescent to perform Reporter gene assay β-galactosidase substrates - MIA PaCa-2

Get tips on using ON-TARGETplus Human PAK1 (5058) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - Caco-2 PAK1

Get tips on using ON-TARGETplus Human SLC7A2 (6542) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - Caco-2 SLC7A2

Get tips on using ON-TARGETplus Human THBS2 siRNA to perform siRNA / miRNA gene silencing Human - Aortic smooth muscle cell TSP-2

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 OxiSelect™ Intracellular ROS Assay Kit (Green Fluorescence) to perform ROS assay cell type - MiaPaCa-2 pancreatic carcinoma

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - MiaPaCa-2 pancreatic carcinoma

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 OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - Capan-2, PANC-1 pancreatic carcinoma