pcDNA3.1 His B
Mammalian vector for expressing proteins with a cleavable N-terminal 6xHis tag. For other reading frames, use pcDNA™3.1/His A or pcDNA™3.1/His C.
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The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible.
PciI is inhibited by nonionic detergents.
Sticky ends from different BsmI sites may not be compatible.
Sticky ends from different PfoI sites may not be compatible.
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.For full activity, add fresh DTT.
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.Sticky ends from different Bpu10I sites may not be compatible.
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
Sticky ends from different Bsu36I sites may not be compatible.
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.