RNA sequencing Human

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1) Lipid

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Bacteria - Gram positive Staphylococcus saprophycitius

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Bacteria - Gram negative Klebsiella pneumoniae

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Bacteria - Gram negative Salmonella enterica

Get tips on using miRCURY LNA™ microRNA Power Labeling Kits to perform Microarray RNA amplification & Labeling - HUVEC Hy3 and Hy5

Get tips on using miRCURY LNA™ microRNA Power Labeling Kits to perform Microarray RNA amplification & Labeling - LNCaP Hy3 and Hy5

Get tips on using MagMAX™ Total Nucleic Acid Isolation Kit to perform RNA isolation / purification Bacteria - Gram positive Mycobacterium tuberculosis

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 Gibco™ DMEM, high glucose to perform Stem cell Differentiation media human umbilical mesenchymal stem cells (hUMSCs) differentiation into osteogenic cells

Get tips on using BrainPhys™ Without Phenol Red to perform Stem cell Differentiation media Differentiation of Human iPSCs into Basal Forebrain cholinergic neurons (BFCN)