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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| + | ==SUMMARY== | ||
| + | #Ran polystyrene wash assays on various concentrations and wash compositions of CPX-inserted cells | ||
| + | #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 | ||
| + | #Grew up individual parts on Mer-GFP system | ||
| + | #Currently working on sending Biobrick I13500 to sequencing | ||
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| + | ==FUTURE PLANS== | ||
| + | #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
- 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 cell fluorescence immunoassay (or western blot) with f2620.b0034.cpx.b0014 transformed cells in M9 media
SUMMARY
- Ran polystyrene wash assays on various concentrations and wash compositions of CPX-inserted cells
- 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
- Grew up individual parts on Mer-GFP system
- Currently working on sending Biobrick I13500 to sequencing
FUTURE PLANS
- 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
