rna-isolation-purification-cells-immortalized-brl-3a

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Get tips on using PureYield™ Plasmid Miniprep System Technical Bulletin to perform Plasmid Isolation Agrobacterium tumefaciens

Products Promega PureYield™ Plasmid Miniprep System Technical Bulletin

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.

RNA siRNA / miRNA gene silencing Rat H9c2 NF-κB RelA (p65)

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Human umbilical cord tissue

Products Sigma-Aldrich CelLytic™ MT Cell Lysis Reagent

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.

RNA siRNA / miRNA gene silencing Rat H9c2 14-3-3 f/Ywhaz

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat mesenchymal stem cells (rMSC)

Products Thermo Fisher Scientific Lipofectamine® 2000 Transfection Reagent

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat schwann cells

Products Sigma-Aldrich X-tremeGENE™ HP DNA Transfection Reagent

Get tips on using TRI Reagent™ Solution to perform Protein isolation Bacteria - Vibrio cholerae

Products Thermo Fisher Scientific TRI Reagent™ Solution

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat aortic smooth muscle cells (rASMC)

Products Promega FuGENE® 6 Transfection Reagent

Get tips on using peqGOLD Cycle‐Pure Kit to perform DNA gel extraction / PCR product purification Product size > 15Kb

Products VWR peqGOLD Cycle‐Pure Kit

Get tips on using QIAquick Gel Extraction Kit to perform DNA gel extraction / PCR product purification Product size < 15Kb

Products Qiagen QIAquick Gel Extraction Kit

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