Difference between revisions of "Part:BBa K200009"

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K200009 short</partinfo>
 
<partinfo>BBa_K200009 short</partinfo>
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This sequence codes for the restriction enzyme DpnII.<br>
 
 
<br><br>
 
<br><br>
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This part encodes a ribosome binding site and DpnII coding region (from [https://parts.igem.org/Part:BBa_K200015 BBa_K200015]).<br>
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<br>
 
===Detail===
 
===Detail===
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This is a Type II restriction enzyme that recognises the sequence GATC.  Its activity can be blocked by dam methylation.
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This is a Type II restriction enzyme that recognises the nucleotide sequence 'GATC.' Its activity is blocked by dam methylation.
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The working temperature is 37°C.
 
The working temperature is 37°C.
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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.
 
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[[Image:II09_DpnII.png]]   
 
[[Image:II09_DpnII.png]]   
 
<br><br>
 
<br><br>
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===Usage and Biology===
 
===Usage and Biology===
  
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Enzyme Properties
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Enzyme Properties can be viewed at the [http://www.neb.com/nebecomm/products/productR0543.asp DpnII Datasheet] from New England Biolabs
  
  
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Activity in NEBuffers:
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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.
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NEBuffer 1: Not Recommended
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NEBuffer 2:  Not Recommended
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NEBuffer 3:  100%
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NEBuffer 4:  Not Recommended
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When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
 
  
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Methylation Sensitivity:
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<span class='h3bb'>Sequence and Features</span>
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dam methylation: Blocked
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dcm methylation: Not sensitive
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CpG methylation: Not sensitive
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More information about: Methylation Sensitivity
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Heat Inactivation:
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===Sequence and Features===
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65°C for 20 minutes
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Survival in a Reaction:
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(+ + +) Suitable for an extended or overnight digestion. Enzyme is active > 8 hours.
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More information about: Extended Digests with Restriction Enzymes
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K200009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K200009 SequenceAndFeatures</partinfo>
  

Latest revision as of 21:38, 19 October 2009

DpnII Composite

This part encodes a ribosome binding site and DpnII coding region (from BBa_K200015).

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.


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


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.


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]