RNA sequencing Rat

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 RNeasy PowerClean Pro Cleanup Kit (50) to perform Removal of contamination in RNA Melanin contamination

Get tips on using ExpressArt FFPE Clear RNAready to perform RNA isolation / purification Tissue - human kidney tissue

Get tips on using illustra RNAspin Mini Kit to perform RNA isolation / purification Cells - immortalized BRL 3A

Get tips on using RNAprep pure Kit (For Plant) to perform RNA isolation / purification Yeast - Neurospora crassa

Get tips on using ISOGEN to perform RNA isolation / purification Cells - immortalized CHO

Get tips on using TRIzol Reagent to perform RNA isolation / purification Bacteria - Gram positive Mycobacterium tuberculosis

Get tips on using TRIzol Reagent to perform RNA isolation / purification Bacteria - Gram positive Clostridum botulinum

Get tips on using TRIzol Reagent to perform RNA isolation / purification Bacteria - Gram positive Clostridium tetani

Get tips on using TRIzol Reagent to perform RNA isolation / purification Bacteria - Gram negative Vibro parahaemolyticus