Protein Expression Eukaryotic cells S. pastorianus

Get tips on using pNAnti-Xyl to perform Protein Expression Prokaryotic cells - S. lividans xylanase

Get tips on using pNAnti-Amy to perform Protein Expression Prokaryotic cells - S. lividans Amylase

Get tips on using pNUF-Amy to perform Protein Expression Prokaryotic cells - S. lividans Amylase

Get tips on using pMJS205 to perform Protein Expression Prokaryotic cells - E. coli S. cerevisiae SOX Erv1p

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 pEGFP-INHα to perform Protein Expression Eukaryotic cells - BHK cells INHα

Get tips on using pJAP2 to perform Protein Expression Eukaryotic cells - HEK293 AT1R

Get tips on using pJAP34 to perform Protein Expression Eukaryotic cells - HEK293 A1R

Get tips on using pJAP37 to perform Protein Expression Eukaryotic cells - HEK293 A1R