Removing unwanted sequence from pUC19
Posted 16 August 2011 - 02:56 AM
Posted 16 August 2011 - 05:35 AM
Posted 16 August 2011 - 09:40 AM
Ignore that. This is a result of sequencing using primers intended for the insert, rather than the backbone plasmid. This is sequence for the backbone plasmid, and it will never sequence "correctly" with those primers. We've been using this plasmid for 10 years with consistent results.
Posted 16 August 2011 - 11:32 AM
If you are using SspI you will remove the promoter for the b-lactamase conferring the AmpR. Otherwise AflIII is in a neutral location not interfering with the origin of replication therefore giving green light.