Site Directed Mutagenesis (SDM) Mouse Deletion 3T3-L1

Get tips on using Accell Mouse Nrep (27528) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 P311/Nrep

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.

Get tips on using TruSeq Stranded mRNA to perform RNA sequencing Mouse - 3T3-L1

Get tips on using TruSeq Stranded Total RNA to perform RNA sequencing Mouse - 3T3-L1

Get tips on using Kdm2b siRNA to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 Fbxl10

Get tips on using Stk11 siRNA to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 Stk11

Get tips on using pcDNA-mC/EBPb to perform CRISPR Mouse - Activation 3T3-L1 C/EBPβ

Get tips on using APC BrdU Flow Kit to perform Cell cycle assay mouse - 3T3-L1

Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Mouse - 3T3-L1 cells