Get tips on using pOPINE to perform Protein expression and purification Bacteria - Escherichia coli medin
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 Mini Kit to perform RNA isolation / purification Bacteria - Gram negative Neisseria gonorrhoeae
Get tips on using 2x Laemmli Sample Buffer to perform Protein isolation Bacteria - Vibrio alginolyticus
Get tips on using TRI Reagent™ Solution to perform Protein isolation Bacteria - Vibrio cholerae
Get tips on using Transcriptor First Strand cDNA Synthesis Kit to perform cDNA synthesis Bacteria
Get tips on using RevertAid First Strand cDNA Synthesis Kit to perform cDNA synthesis Bacteria
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram negative Vibro cholerae
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram negative Legionella pneumophilia
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram negative Borrelia burgdorferi
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment