User:Michael Verhoeven/Notebook/iGEM 2009 M. Verhoeven Parts

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(Project Description/Abstract)
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==Project Description/Abstract==
==Project Description/Abstract==
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* Place short description of project or notes regarding this project
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* Bouyant Bacteria Project (BBP)
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* GVP is a cluster of 14 genes involved in gas vesicle production in ''B. megaterium'' and has been succesfully expressed in ''E.coli''. Not all genes are of critical importance and at least 8 genes are needed. Our aim is to improve the bouyant capacity of the vesicles in ''E.coli''.
==Notes==
==Notes==

Revision as of 10:23, 6 July 2009

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Project Description/Abstract

  • Bouyant Bacteria Project (BBP)
  • GVP is a cluster of 14 genes involved in gas vesicle production in B. megaterium and has been succesfully expressed in E.coli. Not all genes are of critical importance and at least 8 genes are needed. Our aim is to improve the bouyant capacity of the vesicles in E.coli.

Notes

Transformation of E. coli TOP10 with BBa_J23109, BBa_J23100 and BBa_J23106

  • BBa_J23109 iGEM plate 2, 2G
  • BBa_J23100 iGEM plate 1, 18C
  • BBa_J23106 iGEM plate 1, 18O

All dry plasmid DNA in the wells was dissolved in 15ul sterile diH2O, and transfered to 500ul cups.

  • Four cups with 50ul competent E.coli TOP10 cells were thawed on ice, and 2ul of dissolved plasmid was added.
  • After 30 min. on ice, the cups were put in a heat block at 37C for 5 min. and followed by 5 min. on ice.
  • To the cells, 800ul TY medium was added and incubated at 37C for 1 hour.
  • 100ul was plated on TY-Agar-Amp plates and incubated o.n. at 37C.
  • The remainer was centrifuged for 1 min. at full speed and the cells were resuspended in 100ul TY-medium, and plated on TY-Agar-Amp plated. The plates were stored at 37C o.n.

Inoculation of TY-medium with Glycerol Stocks

  • GVP cluster vector BBa_J61035 (2x) in E.coli TOP10
  • Terminator vector ? (2x) in E.coli TOP10
  • 4.5 ml TY-medium with Amp. was inoculated with a colony grown on TY-Agar-Amp. plates o.n.
  • The tubes were put in a waterbath at 37C (150 rpm) and grown o.n.


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