IGEM:MIT/2007/Notebook/2007-8-13

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(New page: ==Agenda== #Check sequencing results of constitutive promoters + CPX #Dilute last night's LCs (F2620.B0034.CPX.B0014) #Miniprep half of last night's LCs and send in for sequencing #Live ce...)
Current revision (11:01, 13 August 2007) (view source)
 
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#Miniprep half of last night's LCs and send in for sequencing
#Miniprep half of last night's LCs and send in for sequencing
#Live cell fluorescence immunoassay (or western blot) with f2620.b0034.cpx.b0014 transformed cells in M9 media
#Live cell fluorescence immunoassay (or western blot) with f2620.b0034.cpx.b0014 transformed cells in M9 media
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 +
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==SUMMARY==
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#Ran polystyrene wash assays on various concentrations and wash compositions of CPX-inserted cells
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#Concluded that overexpression of CPX in more than 10e-5 M AHL is toxic to cell, and most efficient induction happens between 10e-6 and 10e-7 M AHL
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#Grew up individual parts on Mer-GFP system
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#Currently working on sending Biobrick I13500 to sequencing
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 +
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==FUTURE PLANS==
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#Run gel digestion and purify I13500
 +
#Run PCR and purify MerR/MerPT
 +
#Either do two standard assemblies or one 3A assembly on a TK Precut Plasmid
 +
#Send in for sequencing and do mercury assay test

Current revision

Agenda

  1. Check sequencing results of constitutive promoters + CPX
  2. Dilute last night's LCs (F2620.B0034.CPX.B0014)
  3. Miniprep half of last night's LCs and send in for sequencing
  4. Live cell fluorescence immunoassay (or western blot) with f2620.b0034.cpx.b0014 transformed cells in M9 media


SUMMARY

  1. Ran polystyrene wash assays on various concentrations and wash compositions of CPX-inserted cells
  2. Concluded that overexpression of CPX in more than 10e-5 M AHL is toxic to cell, and most efficient induction happens between 10e-6 and 10e-7 M AHL
  3. Grew up individual parts on Mer-GFP system
  4. Currently working on sending Biobrick I13500 to sequencing


FUTURE PLANS

  1. Run gel digestion and purify I13500
  2. Run PCR and purify MerR/MerPT
  3. Either do two standard assemblies or one 3A assembly on a TK Precut Plasmid
  4. Send in for sequencing and do mercury assay test
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