DNA transfection Mammalian cells

Get tips on using CellTiter-Glo® Luminescent Cell Viability Assay to perform Live / Dead assay mammalian cells - BHK-21

Hello everyone! I am currently using different DNA isolation kits to extract DNA from insects. Even though I am able to successfully extract DNA I would like to maximize the yield. Do you have any tips that might help me with that even if the kits are not specifically designed for insect samples?

Get tips on using EZ-10 Spin Column Plasmid DNA Miniprep Kit to perform Plasmid Isolation Cronobacter sakazakii

Get tips on using Pierce™ Cell Surface Protein Isolation Kit to perform Protein isolation Mammalian cells - HUVEC

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 Live/Dead Cell Double Staining Kit to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using Live-Dead cell staining kit (Enzo) to perform Live / Dead assay mammalian cells - human fibroblast tissue