When we isolate plasmid from bacterial culture, usually lb medium is discarded after we pellet down bacterial cells. So my query is what if I directly add TEG buffer to LB medium containing bacterial culture and then pellet the bacterial cells and further process the bacterial culture according to alkaline lysis method for plasmid isolation.
The reason for using TEG is to denature the DNase and make cells permeable to SDS which breaks down the lipids and releases the chromosomal DNA and plasmid DNA into the solution. Chromosomal DNA is bound to membrane fragments and get separated and after neutralization the plasmid is precipitated. LB will reduce the effectiveness of the EDTA allowing DNase activity and the alkalinity of the SDS solution, reducing the efficiency of the lysis, so you can end up with poor quality, poor yield plasmid, contaminated with chromosomal DNA, so it is better not to skip this step.
Because the media components, extracellular microbial primary and secondary metabolites and other impurities will significantly affect your downstream procedures. What is the advantage of maintaining the extracellular components when your desired compounds are intracellular?
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