Team:TU Darmstadt/Materials/PstI

From 2012.igem.org

(Difference between revisions)
(Created page with "<html> <link rel="stylesheet" href="https://2012.igem.org/wiki/index.php?title=Team:TU_Darmstadt/css&action=raw&ctype=text/css" type="text/css" /> <div id="TUD"> <div id="...")
(→PstI)
Line 41: Line 41:
== PstI ==
== PstI ==
=== About ===
=== About ===
-
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.
+
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 5' end overhang.
=== Working conditions ===
=== Working conditions ===
-
The recognition motive of EcoRI is:<sup>[1]</sup>
+
The recognition motive of EcoRI is:
-
  5' GAATTC --> 5'---G     AATTC---3'
+
  5' CTGCAG --> 5'---CTGCA     G---3'
-
  3' CTTAAG --> 3'---CTTAA     G---5'
+
  3' GACGTC --> 3'---G     ACGTC---5'
-
'''Buffer:''' NEBuffer EcoRI
+
'''Buffer:''' NEBuffer 3 + BSA
-
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>
+
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 ===
-
* NEB http://www.neb.com/nebecomm/products/productR3101.asp
+
* NEB http://www.neb.com/nebecomm/products/productR3140.asp
=== References ===
=== References ===
-
[1] http://en.wikipedia.org/wiki/EcoRI
+
[1] http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp
-
 
+
-
[2] http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp
+

Revision as of 18:53, 11 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 5' 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

Retrieved from "http://2012.igem.org/Team:TU_Darmstadt/Materials/PstI"