Team:TU Darmstadt/Materials/PstI

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== PstI ==
== PstI ==
=== About ===
=== About ===
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PstI is a endonucleasic [https://2012.igem.org/Team:TU_Darmstadt/Materials#Restriction_enzymes restriction enzyme] isolated from ''[http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli RY13]''. It creates stick ends with 5' end overhang.
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PstI is a endonucleasic [https://2012.igem.org/Team:TU_Darmstadt/Materials#Restriction_enzymes restriction enzyme] isolated from ''[http://en.wikipedia.org/wiki/Providencia_stuartii Providencia stuartii 164]''. It creates stick ends with 3' end overhang.
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=== Working conditions ===
=== Working conditions ===
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The recognition motive of EcoRI is:<sup>[1]</sup>
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The recognition motive of EcoRI is:
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  5' GAATTC --> 5'---G     AATTC---3'
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  5' CTGCAG --> 5'---CTGCA     G---3'
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  3' CTTAAG --> 3'---CTTAA     G---5'
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  3' GACGTC --> 3'---G     ACGTC---5'
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'''Buffer:''' NEBuffer EcoRI
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'''Buffer:''' NEBuffer 3 + BSA
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EcoRI can exhibit star activity depending on the conditions present in the reaction and needs at least 1bp overhang on its cutting site.<sup>[2]</sup>
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PstI requires at least 1bp overhang on its cutting site (with reduced cleavage efficiency 37%), more than 3bp (98%) are recommended.<sup>[1]</sup>
=== Supplier ===
=== Supplier ===
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* NEB http://www.neb.com/nebecomm/products/productR3101.asp
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* NEB http://www.neb.com/nebecomm/products/productR3140.asp
=== References ===
=== References ===
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[1] http://en.wikipedia.org/wiki/EcoRI
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[1] http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp
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[2] http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp
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Latest revision as of 20:37, 26 September 2012

Contents

PstI

About

PstI is a endonucleasic restriction enzyme isolated from [http://en.wikipedia.org/wiki/Providencia_stuartii Providencia stuartii 164]. It creates stick ends with 3' end overhang.

Working conditions

The recognition motive of EcoRI is: 

5' CTGCAG --> 5'---CTGCA     G---3'
3' GACGTC --> 3'---G     ACGTC---5'

Buffer: NEBuffer 3 + BSA

PstI requires at least 1bp overhang on its cutting site (with reduced cleavage efficiency 37%), more than 3bp (98%) are recommended.[1]

Supplier

  • NEB http://www.neb.com/nebecomm/products/productR3140.asp

References

[1] http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp

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