Protein Expression Eukaryotic cells S. pastorianus

Get tips on using QuickGene RNA Cultured Cell HC Kit S to perform RNA isolation / purification Cells - immortalized SH-SY5Y

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 EMBacY#5 to perform Protein Expression Eukaryotic cells - Hi5 Soluble G protein (Hendra Virus)

Get tips on using pPICZαA to perform Protein Expression Eukaryotic cells - P. pastoris SCF

Get tips on using pPpT4_CalB to perform Protein Expression Eukaryotic cells - P. pastoris Lipase

Get tips on using pPuzzle to perform Protein Expression Eukaryotic cells - P. pastoris GFP

Get tips on using P_GAP to perform Protein Expression Eukaryotic cells - P. pastoris GFP

Get tips on using P_AOX1Syn to perform Protein Expression Eukaryotic cells - P. pastoris GFP

Get tips on using pPICZαB to perform Protein Expression Eukaryotic cells - P. pastoris Laccase

Get tips on using KpTrm1p to perform Protein Expression Eukaryotic cells - P. pastoris phytase