Xba I *DISCONTINUED* - FAQs

I have cloned my gene into my vector and then transformed into TOP10 cells. I then did a plasmid miniprep followed by digestion of the DNA with Xba I. However, the vector is not cutting correctly. Can you help me troubleshoot?

The Xba I cutting site is a Dam-methylation sensitive restriction site. E.coli strains that are dam(+) strains, like TOP10, express the methylating enzyme, Dam. You can try re-transforming into a dam(–) strain, such as INV110. Other dam (and dcm) sensitive restriction sites include the following:
Dam: Bcl I, Cla I, Hph I, Mbo I, Mbo II (Nde II), Taq I, Xba I, BspH I, Nru I
Dcm: Ava II, EcoO 109 I, EcoR II, Sau96 I, ScrF, Stu I, Aat I, Apa I, Ban I, Sfi I