Difference between revisions of "Part:BBa K200009"

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:

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]

@@ Line 13: / Line 13: @@
-===Usage and Biology===
 DpnII is a 4 base cutter, leaving a 4 base overhang.
@@ Line 22: / Line 21: @@
 [[Image:II09_DpnII.png]]
 <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:
+°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>
 <partinfo>BBa_K200009 SequenceAndFeatures</partinfo>