Site Directed Mutagenesis (SDM) Human Deletion MCF-7

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized SKBR3, MDA-MB231 and MCF7

Get tips on using 96-Well Cell Invasion Assay, Basement Membrane to perform Cell migration / Invasion cell type - MCF7

Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized SKBR3, MDA-MB231 and MCF7

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - MCF7

Get tips on using STEAP2 metalloreductase to perform siRNA / miRNA gene silencing Human - PC3 (human prostate cancer cell line) STEAP2

Get tips on using QCM Chemotaxis Cell Migration Assay, 24-well (8 µm), colorimetric to perform Cell migration / Invasion cell type - MCF7

Get tips on using Culture-Insert 4 Well in µ-Dish 35 mm, high to perform Cell migration / Invasion cell type - MCF7

Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) have been greatly used for studies on embryonic development and cell differentiation.iPSCs provide a stable source for either self-renewal or differentiation into suitable cells when cultured in a particular environment. Pluripotent cell culture was originally started by deriving cells from inner cell mass (ICM) from pre-implanted blastocysts, these were called embryonic stem cells. These cells after isolation can be grown on traditional extracellular matrices (like mouse embryonic fibroblasts, MEFs) or feeder-free culture systems. DMEM/F12 has been the most commonly used basal media in the culture of pluripotent cells. These cells are cultured at normal atmospheric oxygen levels, 21%, however, some studies have proposed that 4% oxygen tension may be better for hESC growth. Higher D-glucose concentration (4.2g/l) and osmolarity (320mOsm) that mimics the natural environment of embryonic tissue are optimal for the growth of hESCs. Supplements like N2 and/or B-27, in the presence of growth factors like bFGF, have been shown to increase pluripotency of these cells. bFGF, FGF2 and other ligands of receptor tyrosine kinases like IGF are also required or maintain self-renewal ability of these cells. TGF𝛃1, by its activation of SMAD2/3 signalling, also represses differentiation of iPSCs. Other compounds like ROCK inhibitors reduce blebbing and apoptosis in these cells to maintain their clonogenicity. However, an inhibitor for LIF (leukaemia inhibitory factor, which is one of the pluripotent genes) has an opposing effect. Therefore, it is important to understand the culture conditions and media composition that affect downstream signalling in hESCs or iPSCs that may lead to their differentiation.

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.