protein-expression-and-purification-bacteria-escherichia-coli-siva1

Get tips on using RiboPure™ RNA Purification Kit, bacteria to perform

Get tips on using RiboPure™ RNA Purification Kit, bacteria to perform

The challenge in isolating RNA from S. aureus cells is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads is considered to be the better alternative.

Get tips on using IBI’s Todd-Hewitt Broth to perform Bacterial cell culture media Escherichia coli

Get tips on using ExpiCHO™ Expression System Kit to perform Protein expression and purification Mammalian cells - CHO-K1 SUMO protein

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 Difco™ Brain Heart Infusion Agar, BD to perform Bacterial cell culture media Escherichia coli

Get tips on using EasySelect™ Pichia Expression Kit to perform Protein expression and purification Yeast - Pichia pastoris Chymase

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 AT2R

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 AT1R