Protein Expression Prokaryotic cells S. lividans

Get tips on using pANC209 (xylA) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylA

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 pTip-QC2-gi_21218674 to perform Protein Expression Prokaryotic cells - R. erythropolis putative DNA-binding protein

Get tips on using pDG101 to perform Protein Expression Prokaryotic cells - E. coli mScarlet-I

Get tips on using pFR2 to perform Protein Expression Prokaryotic cells - E. coli ferulic acid

Get tips on using pCB4270V4BU-amy to perform Protein Expression Prokaryotic cells - L. citreum amy

Get tips on using pCB4270BU-amy to perform Protein Expression Prokaryotic cells - L. citreum amy

Get tips on using pET32a- lysostaphin to perform Protein Expression Prokaryotic cells - E. coli lysostaphin

Get tips on using pTXB1-MnP1 to perform Protein Expression Prokaryotic cells - E. coli MnP1

Get tips on using pET3a-hsEH to perform Protein Expression Prokaryotic cells - E. coli hsEH