DNA transfection Mammalian cells

Get tips on using Easy-Spin (DNA free) Total RNA Extraction Kit to perform RNA isolation / purification Tissue - Mouse Cornea

Get tips on using EZ-10 Spin Column Plasmid DNA Miniprep Kit to perform Plasmid Isolation E. coli-S. cerevisiae transconjugate

Get tips on using RealTime-Glo™ MT Cell Viability Assay to perform Live / Dead assay mammalian cells - HEK 293

Get tips on using RealTime-Glo™ MT Cell Viability Assay to perform Live / Dead assay mammalian cells - THP-1

Get tips on using Easy-Spin (DNA free) Total RNA Extraction Kit to perform RNA isolation / purification Bacteria - Gram negative Vibro parahaemolyticus

Get tips on using illustra GFX PCR DNA and Gel Band Purification Kits to perform

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 CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Live / Dead assay mammalian cells - CHO-K1

Get tips on using DNeasy Blood & Tissue Kit to perform DNA isolation / purification Cells - Primary cells HUVEC

Get tips on using illustra tissue and cells genomicPrep Mini Spin Kit to perform DNA isolation / purification Tissue - kidney