We digested pUC57 with the XcmI enzyme, and a non-specific band at 1500bp appeared alongside our intended band (2750bp). To diagnose the issue, we attempted gel purification on both the digested and undigested products, but received the same result. Additionally, we tried heating the digested and undigested products to dissolve secondary plasmid formation, but the same result occurred.
is there anyone with same issue who can help us please?
Hi there,
According to the data from Addgene, pUC57 contains no XcmI site...
Dominique Liger
Dear Professor Liger, Thank you for your response. However, I would like to clarify that we have cloned a linker that contains an XcmI site.
Hi again,
As a control I suggest to perform a single digest with an other enzyme cutting the plasmid at a unique site.
Dominique Liger We also tried that, with KpnI and BamHI there was no extra bond, but with XhoI and XcmI the extra band appears again(sometimes with XhoI and sometimes with XcmI)