Team:Frankfurt/Parts

From 2012.igem.org

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=Parts=
=Parts=
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An important aspect of the iGEM competition is the use and creation of standard  biological parts. Each team will make new parts during iGEM and will place them in the [http://partsregistry.org Registry of Standard Biological Parts]. The iGEM software provides an easy way to present the parts your team has created . The "groupparts" tag will generate a table with all of the parts that your team adds to your team sandbox. Note that if you want to document a part you need to document it on the [http://partsregistry.org Registry], not on your team wiki.
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==BioBrick Assembly==
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The BioBrick plasmid pSB1C3 was sent as a linear fragment with blunt ends. The problem is that we wanted to transform this plasmid into ''E.coli'' to get a higher amount of it. Therefore we made a blunt end ligation to transform the circular plasmid into ''E.coli''. After plasmid preparation we get a higher amount of the plasmid but when we wanted to digest pSB1C3 to assemble the BioBrick parts only a small amount of it had been linearized. <br><br>
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Remember that the goal of proper part documentation is to describe and define a part such that it can be used without a need to refer to the primary literature. The next iGEM team should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for  users who wish to know more.
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So after a few trys we decided to take the BioBrick device with the red  fluorescent protein (RFP). We transformed it into ''E.coli'' and made a plasmid preparation. Then we digest it as well and put it together with our digested BioBrick fragments. After ligation we transformed it again into ''E.coli''. After some colonies were grown we didn't see the expected differences of colonies that take the RFP insert again and colonies that contain our wished BioBrick device. So we put the agar plates into the fridge. After some our we looked for it again and we could see very well the differences between the red and the white colonies. <br><br>
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The control digestion of our expected BioBrick devices shows that we get BioBricks of the following parts:<br>
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1. Protein <br>
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<groupparts>iGEM012 Frankfurt</groupparts>
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*HMG-CoA-Reductase
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*FPP-Synthase
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*bifuntional Cyclase
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*ent-Kaurene Oxidase
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2. Promoter<br>
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*pHXT7
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*pPGK1
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*pPFK1
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3. Terminator<br>
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*tHXT7
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*tPFK2
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*tCYC1<br>
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After we have made a "Midi-Preparation" we also give the BioBricks away for sequencing reaction. The results here show, that the terminator and promoter BioBricks have no insert. So we now have to check if these BioBrick parts really have no insert.<br>
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All four proteins show positive results.<br>
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However we leave all BioBrick parts in the parts registry with the suitable sequences until we have checked our results again.

Revision as of 03:36, 27 September 2012

Team: iGEM Frankfurt - 2012.igem.org

Parts

BioBrick Assembly

The BioBrick plasmid pSB1C3 was sent as a linear fragment with blunt ends. The problem is that we wanted to transform this plasmid into E.coli to get a higher amount of it. Therefore we made a blunt end ligation to transform the circular plasmid into E.coli. After plasmid preparation we get a higher amount of the plasmid but when we wanted to digest pSB1C3 to assemble the BioBrick parts only a small amount of it had been linearized.

So after a few trys we decided to take the BioBrick device with the red fluorescent protein (RFP). We transformed it into E.coli and made a plasmid preparation. Then we digest it as well and put it together with our digested BioBrick fragments. After ligation we transformed it again into E.coli. After some colonies were grown we didn't see the expected differences of colonies that take the RFP insert again and colonies that contain our wished BioBrick device. So we put the agar plates into the fridge. After some our we looked for it again and we could see very well the differences between the red and the white colonies.

The control digestion of our expected BioBrick devices shows that we get BioBricks of the following parts:
1. Protein

  • HMG-CoA-Reductase
  • FPP-Synthase
  • bifuntional Cyclase
  • ent-Kaurene Oxidase

2. Promoter

  • pHXT7
  • pPGK1
  • pPFK1

3. Terminator

  • tHXT7
  • tPFK2
  • tCYC1

After we have made a "Midi-Preparation" we also give the BioBricks away for sequencing reaction. The results here show, that the terminator and promoter BioBricks have no insert. So we now have to check if these BioBrick parts really have no insert.
All four proteins show positive results.
However we leave all BioBrick parts in the parts registry with the suitable sequences until we have checked our results again.

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