Composite

Part:BBa_K200009

Designed by: David Roche, Royah Vaezi   Group: iGEM09_Imperial College London   (2009-08-12)
Revision as of 13:57, 29 September 2009 by Droche (Talk | contribs) (→‎Usage and Biology)

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]



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Parameters
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