Protein expression and purification Mammalian cells HeLa

Get tips on using Total RNA Purification Kit to perform RNA isolation / purification Bacteria - Gram negative Hemophilus influenzae

Get tips on using Genomic DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram negative Salmonella enterica

Get tips on using Genomic DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram positive Staphylococcus aureus

Get tips on using FuGENE® HD Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells HUVEC

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Cardiomyocytes

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 MinElute PCR Purification Kit to perform DNA gel extraction / PCR product purification Product size < 15Kb

Get tips on using Purelink PCR Purification Kit to perform DNA gel extraction / PCR product purification Product size < 15Kb

Get tips on using QIAquick PCR Purification Kit to perform DNA gel extraction / PCR product purification Product size < 15Kb

Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Human - Point mutation HeLa CCNE