siRNA / miRNA gene silencing Rat NRK

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 DYKDDDDK Tag (D6W5B) Rabbit mAb (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #14793 to perform ChIP Anti-bodies FLAG

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - immortalized NK-92MI

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - immortalized NK-92

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized NK-92

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized NK-92MI

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - immortalized NK-92MI

Get tips on using Gibco™ MEM α, Nucleosides to perform Mammalian cell culture media NK-92

Get tips on using AllPrep DNA/RNA Mini Kit to perform RNA isolation / purification Cells - immortalized NK-92