RNA isolation / purification Cells immortalized

Get tips on using QIAprep Spin Miniprep Kit to perform Plasmid Isolation Salmonella enterica serovar Indiana (S. Indiana)

Get tips on using NucleoBond® Xtra Midi / Maxi to perform Plasmid Isolation Shiga toxin-producing E. coli

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 QIAGEN Plasmid Plus 96 Miniprep Kit (4) to perform Plasmid Isolation E. coli DH5α

Get tips on using E.Z.N.A.® Plasmid Mini Kit I, (Q-spin) to perform Plasmid Isolation Actinomyces odontolyticus

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

Get tips on using E.Z.N.A.® Plasmid Mini Kit I, (Q-spin) to perform Plasmid Isolation Salmonella Heidelberg

Get tips on using E.Z.N.A.® Plasmid Mini Kit I, (Q-spin) to perform Plasmid Isolation Bacillus licheniformis

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 GeneJuice® Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human osteoblasts