Throughout my undergraduate education I have been taught that plasmids can't carry very large inserts, but I have never been told why. Any insight would be greatly appreciated.

  • $\begingroup$ Transformation efficiency and the bacteria's ability to maintain the plasmid is reduced as the plasmid size gets larger. Bacterial (300kb) and Yeast (1000-3000kb) Artificial chromosomes can be pretty big, but it is more difficult to get them to work. Also most of what you are working with won't be that big. Inserts usually come from cDNA produced by PCR from mature mRNAs. Without introns, the majority of genes you will see won't be more than a few KB, so plasmids do not need to be that large. $\endgroup$
    – AMR
    Oct 15, 2015 at 3:14


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