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Get tips on using MACSprep™ HLA B/T Cell Isolation Kit, human to perform Cell Isolation HLA B/T

Products Miltenyibiotec MACSprep™ HLA B/T Cell Isolation Kit, human

Get tips on using SmGMTM- 2 Smooth Muscle Cell Growth Medium -2 BulletKitTM to perform Mammalian cell culture media HCASMC

Products Lonza SmGMTM- 2 Smooth Muscle Cell Growth Medium -2 BulletKitTM

Get tips on using SmGMTM- 2 Smooth Muscle Cell Growth Medium -2 BulletKitTM to perform Mammalian cell culture media HCtASMC

Products Lonza SmGMTM- 2 Smooth Muscle Cell Growth Medium -2 BulletKitTM

Get tips on using REPLI-g Cell WGA & WTA Kit (48) to perform Whole Transcriptome Amplification Virus

Products Qiagen REPLI-g Cell WGA & WTA Kit (48)

Get tips on using REPLI-g WTA Single Cell Kit (96) to perform Whole Transcriptome Amplification Virus

Products Qiagen REPLI-g WTA Single Cell Kit (96)

Get tips on using Tissue or Cell Total Protein Extraction Kit to perform Protein isolation Tissue - Lamb muscle tissue

Products Sangon Biotech Tissue or Cell Total Protein Extraction Kit

Get tips on using EasySep™ Human B Cell Enrichment Kit II Without CD43 Depletion to perform Cell Isolation B cell

Products STEMCELL technologies EasySep™ Human B Cell Enrichment Kit II Without CD43 Depletion

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 Human hES cell line H1 (WA01) B2M

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 Human hES cell line H1 (WA01) LIN28

Cell culture media 3D Cell Culture Media Murine colonic organoids

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