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【求助】关于NEB公司Hind3和Ecor1双酶切的技术讨论
大家好， 想就这个双酶切的问题进行讨论一下。我第一次尝试是 两个酶放在一起反应的， 加了两种Bufffer，但是电泳一个条带都没有， 大家有做这方面的经验吗
hsuanzxz 2015-11-09
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Cut 1-2 ug of plasmid DNA with EcoR I + Hind III using NEB 10 x buffer 2.1 in a final volumes of 20-30 ul and incubate at 37 C for 2 hours. Alqueire 3-5 ul of the digestions and run gel to examine the digestion.
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加一种buffer就行了啊，你不会50μl体系各加了5μl吧。。酶的protocal上有写在NEB不同buffer中的酶活力，你选一个都是100%的就行了，NEB官网也有，http://www.neb.sg/tools-and-resources/interactive-tools/double-digest-finder，你这两种酶应该用BUFFER2.1
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