There may be two potential possibilities for this:
- If the dbDNA structure was not formed (e.g., EnClose™ TelN Protelomerase was excluded from the workflow), the sequence of interest (SOI) will be degraded in the enzymatic cleanup step during the restriction enzyme/exonuclease incubation. Confirm that the input plasmid correctly contains the telRL sequences. Include a control reaction with the dbDNA Vector to ensure that the enzymes are working correctly.
- The restriction enzyme used during the enzymatic cleanup cuts within the sequence of interest-containing dbDNA, leaving exposed ends for exonuclease digestion. We recommend using NEBcutter to ensure the processing restriction enzyme recognition sequences only reside outside the SOI.
