RNA sequencing Rat

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary hematopoietic stem cells

Get tips on using DNase Max Kit (50) to perform Removal of contamination in RNA DNA contamination

Get tips on using FastLane Cell Probe Kit (200) to perform RNA isolation / purification Cells - immortalized A549

Get tips on using PolyATtract® mRNA Isolation Systems to perform RNA isolation / purification Yeast - Coprinus cinereus

Get tips on using SensiFAST™ Probe No-ROX One-Step Kit to perform RNA quantification qPCR

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 NucleoSpin® miRNA to perform RNA isolation / purification Cells - primary canine aortic endothelial cells

Get tips on using NucleoSpin® miRNA to perform RNA isolation / purification Cells - primary human mesenchymal stem cells

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Cells - primary human endometrial stromal cells

Get tips on using miRcute miRNA Isolation Kit to perform RNA isolation / purification Cells - immortalized MDA-MB-157