Difference between revisions of "Part:BBa K200009"

(→‎Usage and Biology)
(→‎Detail)
Line 5: Line 5:
 
<br><br>
 
<br><br>
 
===Detail===
 
===Detail===
−
This is a Type II restriction enzyme that recognises the sequence GATC.  Its activity can be blocked by dam methylation.
+
This is a Type II restriction enzyme that recognises the nucleotide sequence 'GATC.' Its activity is blocked by dam methylation.
  
 
The working temperature is 37°C.
 
The working temperature is 37°C.
  
−
As part of the Imperial 2009 iGEM E.ncapsulator project, TaqI was used along with DpnII to cut up the genetic material within the cell, completing the cell death module.  
+
The gene was used alongside with the restriction enzyme TaqI and the methylase Dam by the Imperial iGEM 2009 [http://2009.igem.org/Team:Imperial_College_London <i>The E.ncapsulator</i>] project as part of the genome deletion module.
  
  
Line 21: Line 21:
 
[[Image:II09_DpnII.png]]   
 
[[Image:II09_DpnII.png]]   
 
<br><br>
 
<br><br>
−
 
  
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 14:13, 29 September 2009

DpnII Composite

This sequence codes for the restriction enzyme DpnII.


Detail

This is a Type II restriction enzyme that recognises the nucleotide sequence 'GATC.' Its activity is blocked by dam methylation.

The working temperature is 37°C.

The gene was used alongside with the restriction enzyme TaqI and the methylase Dam by the Imperial iGEM 2009 [http://2009.igem.org/Team:Imperial_College_London The E.ncapsulator] project as part of the genome deletion module.



DpnII is a 4 base cutter, leaving a 4 base overhang.


The cutting site is as shown:

II09 DpnII.png

Usage and Biology

Enzyme Properties can be viewed at the [http://www.neb.com/nebecomm/products/productR0543.asp DpnII Datasheet] from New England Biolabs


Methylation Sensitivity: dam methylation: Blocked dcm methylation: Not sensitive CpG methylation: Not sensitive More information about: Methylation Sensitivity

Heat Inactivation: 65°C for 20 minutes

Survival in a Reaction: (+ + +) Suitable for an extended or overnight digestion. Enzyme is active > 8 hours. More information about: Extended Digests with Restriction Enzymes


Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[http://www.neb.com/nebecomm/products/productR0543.asp Information from NEB]