How are pUC vectors designed for efficient sequencing of insert DNA?

[TytoAlba95]

'The MCSs of many vectors such as the pUC series are flanked by sequences complementary to a universal series of primers, the M13 forward and reverse primers. These priming sites are oriented such that extension of the primers annealed to these sites allows sequencing of both ends of an insert DNA in the MCS. In this fashion, one set of universal primers can be used to sequence any insert DNA regardless of which site the DNA was inserted at within the MCS'I'm unable to imagine how the insert with the primer can be separated from the ds plasmid for sequencing.

 

[Ygggdrasil]

Primers for sequencing are usually synthesized chemically. They are mixed with the plasmid along with other reagents to allow the sequencing reaction to occur. The primers bind to sequences on the plasmid.

 

[TytoAlba95]

Thank you.