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 Mouse Chitinase 3-like 1 Quantikine ELISA Kit to perform ELISA Mouse - Chitinase-3-Like Protein-1 (CHI3L1) or YKL-40
DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.
The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.
Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - Capan-2, PANC-1 pancreatic carcinoma
Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - PLHC-1 poeciliopsis lucida hepatocellular carcinoma
Get tips on using Monoclonal Mouse Anti-Human Cytokeratin (Concentrate) Clone AE1/AE3 to perform Immunohistochemistry Mouse - Cytokeratin
Get tips on using Purified Rat Anti-Mouse CD24 Clone M1/69 (RUO) to perform Immunohistochemistry Mouse - CD24
Get tips on using Purified Rat Anti-Mouse CD24 Clone M1/69 (RUO) to perform Immunohistochemistry Mouse - CD24
Get tips on using Mouse GFR alpha-3/GDNF R alpha-3 Antibody to perform Immunohistochemistry Mouse - Gfrα3
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