Difference between revisions of "Part:BBa K200009"
(→‎Usage and Biology)
Line 13: Line 13:
  
  
−
===Usage and Biology===
 
  
 
DpnII is a 4 base cutter, leaving a 4 base overhang.
 
DpnII is a 4 base cutter, leaving a 4 base overhang.
Line 22: Line 21:
 
[[Image:II09_DpnII.png]]   
 
[[Image:II09_DpnII.png]]   
 
<br><br>
 
<br><br>
 +
 +
 +
===Usage and Biology===
 +
 +
Enzyme Properties
 +
 +
 +
Activity in NEBuffers:
 +
NEBuffer 1:  Not Recommended
 +
NEBuffer 2:  Not Recommended
 +
NEBuffer 3:  100%
 +
NEBuffer 4:  Not Recommended
 +
 +
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
 +
 +
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
 +
 +
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K200009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K200009 SequenceAndFeatures</partinfo>

Revision as of 13:57, 29 September 2009

DpnII Composite

This sequence codes for the restriction enzyme DpnII.


Detail

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

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.



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


Activity in NEBuffers: NEBuffer 1: Not Recommended NEBuffer 2: Not Recommended NEBuffer 3: 100% NEBuffer 4: Not Recommended

When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

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


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]