Melbourne/Lab GV Notebook

From 2007.igem.org

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[[Melbourne/Lab Notebook|<Return to Lab notebook>]]  [[Melbourne|  <team home page>]]
[[Melbourne/Lab Notebook|<Return to Lab notebook>]]  [[Melbourne|  <team home page>]]
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*Image:Image-melbourne-Development path gv.jpg|850px|Development of BBa_I750016]]
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*[[Image:Image-melbourne-Development path gv.jpg|850px|Development of BBa_I750016]]
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*[[Melbourne/Lab Notebook gv 0|Synopsis]]
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*Synopsis:
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The polycistronic gas vesicle gene was obtained in an expression plasmid. We followed the [[http://partsregistry.org/Help:Standardization|normal process]] for converting it to a biobrick. The only complicating factor should have been the large size of the part (11Kbp approx). To help with this we used the stratagene XL Quickchange kit for site dirrected mutagenesis, and XL-10 gold ultra competent cells.
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Things were going swimmingly until we tried to digest the biobrick prefix & suffix and ligate it into BBa_J61035 which carries an RBS with agentamycin handle. The digestion, purification, and ligation processes present a large number of options but very few points of intermediate observation.
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=Steps:=
=Steps:=
*[[Melbourne/Lab Notebook gv 1|Create Glycerol stocks of gas vesicle plasmids and cells supplied by Maura Cannon]]
*[[Melbourne/Lab Notebook gv 1|Create Glycerol stocks of gas vesicle plasmids and cells supplied by Maura Cannon]]

Revision as of 07:59, 25 October 2007

<Return to Lab notebook> <team home page>

  • Development of BBa_I750016
  • Synopsis:

The polycistronic gas vesicle gene was obtained in an expression plasmid. We followed the [process] for converting it to a biobrick. The only complicating factor should have been the large size of the part (11Kbp approx). To help with this we used the stratagene XL Quickchange kit for site dirrected mutagenesis, and XL-10 gold ultra competent cells. Things were going swimmingly until we tried to digest the biobrick prefix & suffix and ligate it into BBa_J61035 which carries an RBS with agentamycin handle. The digestion, purification, and ligation processes present a large number of options but very few points of intermediate observation.

Steps: