Skill, knowledge, determination, acumen and connections – along with a little luck and serendipity – all can play a part. The 1-base overhangs produced by PflFI may be hard to ligate.The successful launch and longevity of a startup is often the result of several factors. Sticky ends from different BstXI sites may not be compatible. This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.Ĭ C A N N N N N N T G G G G T N N N N N N A C C Sticky ends from different PfoI sites may not be compatible. Sticky ends from different Bsu36I sites may not be compatible. SmaI can be used at 37☌ for brief incubations. Sticky ends from different BstEII sites may not be compatible.īstEII is typically used at 60☌, but is 50% active at 37☌.Ĭleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. Sticky ends from different PpuMI sites may not be compatible.Įfficient cleavage requires at least two copies of the NarI recognition sequence.Ĭleavage may be enhanced when more than one copy of the BbeI recognition sequence is present. Sticky ends from different EcoO109I sites may not be compatible. PshAI quickly loses activity at 37☌, but can be used at 25☌ for long incubations. Sticky ends from different XcmI sites may not be compatible.īsrGI is typically used at 37☌, but is even more active at 60☌.īssHII is typically used at 50☌, but is 75% active at 37☌. The 1-base overhangs produced by XcmI may be hard to ligate. Prolonged incubation with NdeI may lead to removal of additional nucleotides.ĮcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.Īfter cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.Ĭ C A N N N N N N N N N T G G G G T N N N N N N N N N A C C PaeR7I does not recognize the sequence CTCTCGAG. Sticky ends from different AhdI sites may not be compatible.Įfficient cleavage requires at least two copies of the SacII recognition sequence.
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