After alkaline lysis, there is an additional isopropanol precipitation step to decrease the amount of lysate before DNA is bound to the QIAGEN-tip.

The recommended conditions below are suitable for QIAGEN-tip 100 or QIAGEN-tip 500, and use centrifugation to clear lysates rather than QIAfilter Cartridges, due to the large culture volumes. Very low-copy plasmids and cosmids of less than 10 copies per cell often require large culture volumes to yield significant amounts of DNA. The high-copy plasmids listed here contain mutated versions of this origin. * The pMB1 origin of replication is closely related to that of ColE1 and falls in the same incompatibility group.

Origins of replication and copy numbers of various plasmids and cosmids A high-copy plasmid should yield between 3-5 ug DNA per 1 ml LB culture, while a low-copy plasmid will yield between 0.2-1 ug DNA per ml of LB culture. A way to determine experimentally if the copy number of your plasmid is high or low is to perform a miniprep. This table can also be found online at the QIAGEN Plasmid Resource Center in the section ' Growth of bacterial cultures Plasmid Copy Number'. Transfection, cloning, sequencing, capillary sequencing, etc.įind out which origin of replication your plasmid contains, and look at the table below for classification into high-copy or low-copy types.

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