【求助】3’末端切平
丁香园论坛
1975
请问有谁做过3’末端切平啊？我要做平末端连接，先将目的片段通过限制酶进行酶切，可两端产生的都是3'突出末端，不知道该用那种酶进行切平比较好，看见有好多都是补平的体系，现不知道具体用哪个公司的那种酶效果会比较好，是个新手，谢谢了！
neb不错，值得推荐。
若水无涯 您好！
您如果要做3‘端的切除，NEB有以下几种酶建议您可以考虑：
DNA Polymerase I, Large (Klenow) Fragment and T4 DNA Polymerase (NEB# M0203) are the best choices for chewing back 3' overhangs and filling in 5' overhangs (3' recessed ends).DNA Polymerase I, Large (Klenow) Fragment can be used at 25°C or room temperature, but T4 DNA Polymerase must be used at 12°C due to its robust exonuclease. Both work well in a wide variety of buffers. Vent DNA Polymerase (NEB# M0254) and Deep Vent DNA Polymerase (NEB# M0258) can also be used but ThermoPol buffer must be used, the reaction temperature is high, and the enzyme cannot be heat inactivated. Mung Bean Nuclease (NEB# M0250) will chew back 3' overhangs but the strong exonuclease activity combined with the lack of polymerase activity yield a lower percentage of blunt ends.
祝您试验顺利！
您如果要做3‘端的切除，NEB有以下几种酶建议您可以考虑：
DNA Polymerase I, Large (Klenow) Fragment and T4 DNA Polymerase (NEB# M0203) are the best choices for chewing back 3' overhangs and filling in 5' overhangs (3' recessed ends).DNA Polymerase I, Large (Klenow) Fragment can be used at 25°C or room temperature, but T4 DNA Polymerase must be used at 12°C due to its robust exonuclease. Both work well in a wide variety of buffers. Vent DNA Polymerase (NEB# M0254) and Deep Vent DNA Polymerase (NEB# M0258) can also be used but ThermoPol buffer must be used, the reaction temperature is high, and the enzyme cannot be heat inactivated. Mung Bean Nuclease (NEB# M0250) will chew back 3' overhangs but the strong exonuclease activity combined with the lack of polymerase activity yield a lower percentage of blunt ends.
祝您试验顺利！
本文由丁香园论坛提供，想了解更多有用的、有意思的前沿资讯以及酷炫的实验方法的你，都可以成为师兄的好伙伴
师兄微信号：shixiongcoming
