DNA End Repair Mix - FAQs

查看更多产品信息 DNA End Repair Mix - FAQs (A14321)

2 个常见问题解答

进行限制性内切酶克隆除了连接酶以外还需要什么酶？

对载体进行去磷酸化以降低背景时可以使用：
热失活的碱性磷酸酶，或
牛小肠碱性磷酸酶（提供1 unit/μL或20 units/μL)规格)

如果需要获得磷酸化的DNA平末端（“补平”末端），您可以使用：
可以用DNA End Repair Mix或T4 DNA 聚合酶或E. coli DNA聚合酶Klenow片段（大片段）来产生平末端，因为它们有5’ →3’DNA聚合酶活性（补齐5’突出端）和3’→5’外切酶活性（切除3’突出端）。

What other enzymes besides ligase may I need to perform restriction cloning?

Dephosphorylating the vector to decrease background can be achieved with:
Heat Inactivated Alkaline Phosphatase, or Calf Intestinal Alkalline Phosphatase (available as 1 unit/µL or 20 units/µL)

If you need to create blunt phosphorylated DNA ends (“polishing” the ends), you can use:
DNA End Repair Mix or T4 DNA polymerase or Klenow Fragment (large fragment) of E. coli DNA polymerase to generate blunt ends due to their 5' to 3' DNA polymerase activity (filling-in of 5' overhangs) and 3' to 5' exonuclease activity (chewing back of 3' overhangs).