pBluescript II SK(-)
Standard cloning vector (phagemid excised from lambda ZAPII). The f1 (–) orientation allows rescue of antisense strand ssDNA. pBluescript II KS(–) has a reversed MCS.
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Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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BsaHI is typically used at 37°C, but is even more active at 60°C. |
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Efficient cleavage requires at least two copies of the NmeAIII recognition sequence. Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°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. |
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Sticky ends from different AlwNI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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Sticky ends from different BtgZI sites may not be compatible.After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.BtgZI is typically used at 60°C, but is 75% active at 37°C. |
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Sticky ends from different DraIII sites may not be compatible. |
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Sticky ends from different EcoO109I sites may not be compatible. |
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ApaI can be used between 25°C and 37°C. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.Sticky ends from different AccI sites may not be compatible. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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SmaI can be used at 37°C for brief incubations. |
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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different BtgI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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Sticky ends from different BstXI sites may not be compatible. |
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Sticky ends from different BspQI sites may not be compatible. |
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Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot. |
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Sticky ends from different AflIII sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
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