IGEM06 DNA Distribution Issues

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|On transforming, this biobrick persistently results in segregated colonies, some show the expected green fluorescence but others show surprisingly red fluorescence. IPTG does not have an effect. This has been our experience
|On transforming, this biobrick persistently results in segregated colonies, some show the expected green fluorescence but others show surprisingly red fluorescence. IPTG does not have an effect. This has been our experience
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|Imperial
 
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|<bbpart>BBa_C0060</bbpart>
 
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'''and all other bricks containing aiiA'''
 
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|pSB1A2
 
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|DH5a, Rosetta
 
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|iGEM2006 DNA-1
 
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| We have had persistent issues with our cells being unable to express this protein (AHL-Lactonase). Upon sequencing our constructs it appears that the DNA we received from the registry is an empty vector with no aiiA gene present. Our sequencing results can be seen in this report.
 
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It is very important that these results are checked and verrified as there are 15 avalible (non-working) parts which rely on this gene and 74+ parts which have been designed/built but not submitted to the registry.
 
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'''List of potentioally affected parts'''
 
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*<bbpart>BBa_C0060</bbpart>
 
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*<bbpart>BBa_C0260</bbpart>
 
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*<bbpart>BBa_I0403</bbpart>
 
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*<bbpart>BBa_I0460</bbpart>
 
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*<bbpart>BBa_I13207</bbpart>
 
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*<bbpart>BBa_I4200</bbpart>
 
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*<bbpart>BBa_I4201</bbpart>
 
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*<bbpart>BBa_I4202</bbpart>
 
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*<bbpart>BBa_I4203</bbpart>
 
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*<bbpart>BBa_I4204</bbpart>
 
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*<bbpart>BBa_S01007</bbpart>
 
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*<bbpart>BBa_S01656</bbpart>
 
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*<bbpart>BBa_S01819</bbpart>
 
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*<bbpart>BBa_S01897</bbpart>
 
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*<bbpart>BBa_S01984</bbpart>
 
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Latest revision as of 13:37, 21 October 2006

This page is to keep a record of any troubles that you have when trying to successfully transform parts from either of the two iGEM 2006 DNA Distribution plates (See Biobrick Delivery page for delivery details)

Please include:

  • Biobrick part number
  • Plasmid
  • Cell strain used
  • Plate (iGEM 2006 DNA-1 or DNA-2) and well number


Team Part number Plasmid Cell strain Distribution plate Comments
Example BBa_xxxx pSB1A2 TOP10 iGEM06 DNA-1 we tried 10 times!!
PennState BBa_R0051 pSB1A2, pSB2K3 DH5a iGEM06/05 DNA-1 neither amp plate seems to work
Davidson BBa_B0015 pSB1AK3 JM109 iGEM06 DNA-1 no insert at all; had to use stocks from last year; pSB1A2
Berkeley BBa_P1000 (CmR) pSB1AC3 TG1 iGEM06 DNA-2 Austin says it should be a pSB2K* plasmid; no CmR colonies generated upon transformation
Berkeley BBa_P1001 (tet) pSB1AT3 TG1 iGEM06 DNA-2 Austin says it should be a pSB2K* plasmid; no tetR colonies generated upon transformation
Ljubljana, Slovenia BBa_P1010 (tet) pSB1AT3 DB3.1 iGEM06 DNA-2 (23P) Plasmid pSB1AT3 should have tet resistance, but instead it has Kan resistance (so it should be labeled pSB1AK3).
Cambridge BBa_J04430 pSB1A2 DH5-a; XL-1 Blue; MG-1655; MC1000 iGEM06 DNA-2 (1L) On transforming, this biobrick persistently results in segregated colonies, some show the expected green fluorescence but others show surprisingly red fluorescence. IPTG does not have an effect. This has been our experience


If we see the same Biobrick parts giving multiple teams trouble it will give us a better sense of what we need to fix.

Note: Please take a look and see if your team has been able to successfully transform any of the parts on this list. You might want to contact the team that was having trouble and let them know that you were able to successfully transform that particular part. All of the distribution plates were done in the same method at the same time so they should be the same, but that isn't always the case.

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