"Xba I -""DISCONTINUED""" - FAQs

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2 个常见问题解答

我克隆了我的基因并转化进TOP10细胞中。然后我进行了质粒小抽并对质粒进行Xba I酶切。但是，载体没有被正确切开。你能帮我解决这一问题吗？

XbaI酶切位点是Dam甲基化敏感的酶切位点。dam(+)的E. Coli菌株，如TOP10，表达甲基化酶Dam。您可以尝试重新转化一种dam(-)菌株，例如INV110。下面列出了其它对dam (和dcm)敏感的限制性内切酶位点：

•Dam敏感：Bcl I, Cla I, Hph I, Mbo I, Mbo II (Nde II), Taqα I, Xba I, BspH I, Nru I
•Dcm敏感：Ava II, EcoO 109 I, EcoR II, Sau96 I, ScrF, Stu I, Aat I, Apa I, Ban I, Sfi I

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