Get tips on using OxiSelect™ Comet Assay Kit (3-Well Slides) to perform DNA Damage Assay HEK 293T
Get tips on using OxiSelect™ Comet Assay Kit (3-Well Slides) to perform DNA Damage Assay U-87 MG
Get tips on using QuantiFluor® RNA System to perform RNA quantification Fuorimetric - human plasma
Get tips on using Target sequence1: Seq1-5'-GACTTCACGGGTGGTGTTTCT-3' to perform shRNA gene silencing Human - HEK 293T CAPN5- (Calpains) cationic lipid based
Get tips on using MEK-3 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - BMDMs MEK-3
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 Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - human blood
Get tips on using Qubit RNA HS Assay Kit to perform RNA quantification Fuorimetric - human plasma
Get tips on using Agilent RNA 6000 Pico Kit to perform RNA quantification Fuorimetric - human saliva
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