PCR Quantitative real-time PCR

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary bovine coronary artery smooth muscle cells

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - immortalized PC-3

Get tips on using TriPure Isolation Reagent to perform RNA isolation / purification Cells - primary human pulmonary artery endothelial cells

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - primary human cardiac fibroblasts

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - primary human bronchial epithelial 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.

Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Cells - primary human umbilical vein endothelial cells

Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Cells - primary human dermal microvascular endothelial cells

Get tips on using FuGENE® HD Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines PC12

Get tips on using Timp2 siRNA to perform siRNA / miRNA gene silencing Mouse - BV2 TIMP-2