rna-isolation-purification-tissue-rat-sublingual-glands

Get tips on using NucleoSpin® RNA to perform RNA isolation / purification Cells - immortalized A549

Get tips on using NucleoSpin® RNA to perform RNA isolation / purification Cells - immortalized OVCAR3

Get tips on using RNAsimple Total RNA Kit to perform RNA isolation / purification Cells - immortalized H1299

Get tips on using Qproteome FFPE Tissue Kit (20) to perform Protein isolation Tissue - Human tissue C-MFPE samples

Get tips on using illustra™ RNAspin Mini Isolation Kit to perform RNA isolation / purification Cells - immortalized DH82

Get tips on using illustra™ RNAspin Mini Isolation Kit to perform RNA isolation / purification Cells - immortalized Vero

Get tips on using SENSE mRNA-Seq Library Prep Kit V2 to perform RNA sequencing Rat - Hippocampal tissue

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

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.

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.