Ligations

From 2006.igem.org

(Difference between revisions)
Jump to: navigation, search
 
(2 intermediate revisions not shown)
Line 11: Line 11:
<font color="red">'''KanB'''</font color>
<font color="red">'''KanB'''</font color>
-
*Kan backwards is the only gene that we are completely positive has not been cloned into a plasmid. Further testing is needed to see if it has indeed been cloned.
+
*Kan backwards is the only gene that we are completely positive has not been cloned into a plasmid in the correct orientation.  
-
 
+
=RE and Hix=
=RE and Hix=
We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for  part Part BBa_J3101 [http://partsregistry.org/Part:BBa_J3101] for further details.  
We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for  part Part BBa_J3101 [http://partsregistry.org/Part:BBa_J3101] for further details.  
Line 22: Line 21:
=Cloning Parts of the Constructs=
=Cloning Parts of the Constructs=
-
The underlined portion of each construct represents parts that have been successfully ligated and cloned:
 
-
 
-
pAra-RBS-Hix- <u>TetF-Hix</u>-<u>DFT-RE</u>
 
-
 
-
pAra-Hix-RBS-<u>TetF-Hix</u>-<u>DFT-RE</u>
 
-
 
-
pAra-RBS-Hix-<u>TetB-Hix</u>-<u>DFT-RE</u>
 
-
pAra-Hix- TetB- SBR-Hix-<u>DFT-RE</u>
+
Please see the [[Construction Intermediates]] page for a list of the ligated parts and their part numbers.
-
*SBR= backwards RBS
+

Latest revision as of 15:08, 2 August 2006

Cloning

We have been able to successfully clone the following antibiotic pancakes

  • Hin
  • Hin with LVA degredation tag
  • Cmr- forward
  • Kan-forward
  • Tet-forward
  • Tet-backwards
  • Cmr-backwards


KanB

  • Kan backwards is the only gene that we are completely positive has not been cloned into a plasmid in the correct orientation.

RE and Hix

We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for part Part BBa_J3101 [http://partsregistry.org/Part:BBa_J3101] for further details.

  • As per the [http://2006.igem.org/Davidson_Assembly_Plan| assembly plan] we will be attaching the RE downstream of the double terminator BBa_B0015 [http://partsregistry.org/Part:BBa_B0015]
  • We also have two cultures with the correct sequences for Hix C.
  • We have sent a "Master Plate" with cultures of all of the things that we have cloned to the Mammoths.

Cloning Parts of the Constructs

Please see the Construction Intermediates page for a list of the ligated parts and their part numbers.

Personal tools
Past/present/future years