rna-isolation-purification-tissue-mouse-lung

Get tips on using QuickLyse Miniprep Kit (250) to perform Plasmid Isolation Enterobacteriaceae

Get tips on using MEBMTM Mammary Epithelial Cell Growth Basal Medium to perform 3D Cell Culture Media Mouse primary breast cancer ephitelial cells-Mammospheres

Get tips on using GeneArt™ Site-Directed Mutagenesis PLUS System to perform Site Directed Mutagenesis (SDM) Mouse - Point mutation L929 SigmaR1 gene (σ1)

Get tips on using pSUPER.retro.neo+gfp vector- Syn G (exon 3) siRNA to perform shRNA gene silencing Mouse - RGC-5 Syn G (Exon 3)

Get tips on using FITC Annexin V Apoptosis Detection Kit I (RUO) to perform Apoptosis assay cell type - T-cells Mouse (CD4+ and CD8+)

Get tips on using Silencer® Select Negative Control No 1 siRNA to perform siRNA / miRNA gene silencing Mouse - siRNA negative control polymer / lipid

Get tips on using Anti-Human CD56 (NCAM) APC-eFluor® 780 to perform Flowcytometry CD56 (NCAM) - Mouse / IgG1, kappa Human APC-eFluor 780

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 PE Annexin V Apoptosis Detection Kit with 7-AAD to perform Apoptosis assay cell type - T-cells Mouse (OT-I)

Get tips on using CD279 (PD-1) Monoclonal Antibody (RMP1-30), FITC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD279/PD-1