Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines Huh7
Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HepG2
Get tips on using EZ-Magna ChIP™ A/G Chromatin Immunoprecipitation Kit to perform ChIP Human - Fibroblast cell lines
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 RNeasy Plus Mini Kit to perform RNA isolation / purification Tissue - Mouse Adrenal glands
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Tissue - Mouse Spinal cord
Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Tissue - Mouse Artery / aorta
Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Tissue - Mouse Mammary glands
Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Tissue - mouse aorta tissue
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Tissue - mouse pituitary tissue
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment