Protein isolation Mammalian cells

Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCAEC

Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCtASMC

Get tips on using Human MesoEndo Cell Growth Medium to perform Mammalian cell culture media HCtAEC

Get tips on using Endothelial Cell Growth Medium MV to perform Mammalian cell culture media BAOEC

Get tips on using EpiTect MSP Kit to perform PCR Methylation specific PCR - Mammalian DNA

Get tips on using FastLane Cell Multiplex Kit (200) to perform PCR Multiplex PCR - Mammalian DNA

Get tips on using QIAGEN Multiplex PCR Kit to perform PCR Multiplex PCR - Mammalian DNA

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.

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase