Get tips on using CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) to perform Cell cytotoxicity / Proliferation assay cell type - Hep G2
Hello Iam a phd student in pharmacy and i want to know if this technology is suitable to knockout or silencing part of the gas5 gene in BV2 cells please
Get tips on using APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU to perform DNA Damage Assay HeLa
Get tips on using Corning® 500 mL MEM (Minimum Essential Medium) Alpha Medium to perform Stem cell culture media Cord blood-derived endothelial cells(hCBiPS2)
Get tips on using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - HepG2 FHIT
Get tips on using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - Hep3B SFRP3
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 Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy to perform ROS assay cell type - H9c2 rat cardiomyocytes
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.
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