How can I increase plasmid DNA yield?
Many things affect plasmid yield. For plasmids in E. coli strains, there are two simple things that can increase plasmid yield.
Increase culture volume
• ost plasmid prep sysM tems can process a range of volumes. For example, the PureYield™ Miniprep System can process 600 μl to 3 ml of culture. tains more NaCl, can significantly increase yields, provided that the biomass is within the acceptable range. • ulture incubation times: C Low-density bacterial cultures yield relatively low amounts of DNA. Overgrown cultures produce suboptimal yields and excessive chromosomal DNA contamination. Do not use cultures grown longer than 18–20 hours. • or 1–100 ml of culture: Pick F an isolated colony from a freshly streaked plate, and inoculate LB medium [+ antibiotic(s)]. Incubate with shaking for 8–16 hours at 37 °C before harvesting.
Note: Exceeding the recommended culture volume can result in decreased yields because the increased biomass can lead to insufficient lysis and lysate clearing problems.
Optimize culture conditions
• edia: We recommend growM ing cells in 1X Luria-Bertani (LB) medium. The use of rich medium, such as 2X YT, CircleGROW, Terrific broth, or LB-Miller medium, which con-
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