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Get tips on using QIAseq FX Single Cell RNA Library Kit (96) to perform Whole Transcriptome Amplification Virus

Products Qiagen QIAseq FX Single Cell RNA Library Kit (96)

Get tips on using EGMTM -2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKitTM to perform Stem cell Differentiation media hMSCs differentiation into pericytes

Products Lonza EGMTM -2 MV Microvascular Endothelial Cell Growth Medium-2 BulletKitTM

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) OCT4-PG1

DNA DNA isolation / purification Cells Immortalized cell lines Loucy

DNA DNA isolation / purification Cells Immortalized cell lines MKN45

RNA RNA isolation / purification Supernatant from cell cultures

Get tips on using Flp-In™ T-REx™ 293 Cell Line to perform Protein expression and purification Mammalian cells - HeLa ChaC1

Products Thermo Fisher Scientific Flp-In™ T-REx™ 293 Cell Line

Cell culture media 3D Cell Culture Media PLC/PRF/5 cells-spheres

Cell culture media 3D Cell Culture Media hiPSC-derived lung organoids

Cell culture media 3D Cell Culture Media Human small intestinal organoids

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