Get tips on using siRNA Wee1 to perform siRNA / miRNA gene silencing Human - SKOV-3 Wee-1
Get tips on using Ccl2 siRNA to perform siRNA / miRNA gene silencing Rat - Neuronal cells MCP-1
Get tips on using Hs_IFI16_1 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - TF‐1 IFI16
Get tips on using pSilencer 2.1-U6 Hygro to perform shRNA gene silencing Human - SHSY5Y Beclin 1
Get tips on using AChE shRNA Plasmids (h) to perform shRNA gene silencing Human - TF‐1 AChE
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 QIAamp Fast DNA Tissue Kit to perform DNA isolation / purification Tissue - colon
Get tips on using QIAamp DNA FFPE Tissue Kit to perform DNA isolation / purification Tissue - heart
Get tips on using QIAamp DNA FFPE Tissue Kit to perform DNA isolation / purification Tissue - bone
Get tips on using AllPrep DNA/RNA Mini Kit to perform DNA isolation / purification Tissue - eye
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