Microarray Mice Cochlea

Get tips on using Gibco™ RPMI 1640 Medium to perform Stem cell Differentiation media hiPSCs differentiation into Microglial-like cells

Get tips on using Gibco™Neurobasal™ Medium to perform Stem cell Differentiation media hiPSCs differentiation into Microglial-like cells

Get tips on using RNeasy Lipid Tissue Mini Kit to perform RNA isolation / purification Cells - primary human brain microvascular endothelial cells

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - mouse microglia

Get tips on using VWR Life Science RiboZol™ RNA Extraction Reagent to perform RNA isolation / purification Cells - primary rat brain microvascular endothelial cells

Get tips on using CytoSelect™ 24-Well Wound Healing Assay to perform Wound healing assay cell type - human gHMVEC (glioma human microvascular endothelial cells)

Get tips on using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® to perform RNA sequencing Mouse - Microglia

Get tips on using EZViable™ Calcein AM Cell Viability Assay Kit (Fluorometric) to perform Live / Dead assay mammalian cells - rat brain microvascular endothelial cells

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.

A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.