rna-isolation-purification-cells-immortalized-mda-mb-231

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 Xfect™ Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human pulmonary artery smooth muscle cells (HPASMC)

Get tips on using peqGOLD Cycle‐Pure Kit to perform DNA gel extraction / PCR product purification Product size > 15Kb

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

Get tips on using EasySelect™ Pichia Expression Kit to perform Protein expression and purification Yeast - Pichia pastoris Chymase

Get tips on using Strep-Tactin Superflow Plus (10 ml) to perform Protein tag Purification of Strep-tagged proteins

Get tips on using Ni-NTA Fast Start Kit (6) to perform Protein tag Purification of His-tagged proteins

Get tips on using Quick Amp Labeling Kit-one color to perform RNA amplification & labeling Mammalian - RNA, Mice Cochlaea Cyanine CTP

Get tips on using MagniSort™ Human pan-Monocyte Enrichment Kit to perform Cell Isolation Monocyte

Get tips on using CelLytic™ Y Plus Kit to perform Protein isolation Yeast - Candida boidinii