IGEM:MIT/2006/Notebook/2006-7-14: Difference between revisions
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We got many, many transformants hopefully carrying SAMT connected to B0015 and BSMT connected to B0015. We made 4 liquid cultures from each type of colony. We also made two liquid cultures from the glycerol stock of cells containing the ribosome binding sites B0030 and B0032. I plan to come in tomorrow to miniprep each of these 10 liquid cultures for eventual sequencing (in the case of the SAMT.B0015 and BSMT.B0015 cultures) and ligation (for each culture). | We got many, many transformants hopefully carrying SAMT connected to B0015 and BSMT connected to B0015. We made 4 liquid cultures from each type of colony. We also made two liquid cultures from the glycerol stock of cells containing the ribosome binding sites B0030 and B0032. I plan to come in tomorrow to miniprep each of these 10 liquid cultures for eventual sequencing (in the case of the SAMT.B0015 and BSMT.B0015 cultures) and ligation (for each culture). | ||
==GC== |
Latest revision as of 07:21, 17 July 2006
TO DO
Andre: re-do SPP PCR (followed by PCR cleanup & gel); find out about exponential phase promoter (make liquid culture for miniprep if possible), research other smells
Stephen: Site-directed mutagenesis of ATF1 + gel; Make indole knockout cells competent (to be followed by transforming Dudareva plasmid); re-do SPP PCR (followed by PCR cleanup & gel), make liquid culture of coding region + terminator transformants, make RBS cultures
Veena: GC on different BSMT concentrations and indole knockout, order primers for BAT2, THI3, pCHBA
Indole Knockout Strain
The indole knockout strain was made competent using the following protocol:
http://openwetware.org/wiki/Preparing_chemically_competent_cells
Site-directed mutagenesis for ATF1
Redone using the Strategene manual available here: http://openwetware.org/wiki/Endy:Site-directed_Mutagenesis
We got the primers ready for mutagenesis using the following protocol:
http://openwetware.org/wiki/PNK_Treatment_of_DNA_Ends
Liquid cultures
We got many, many transformants hopefully carrying SAMT connected to B0015 and BSMT connected to B0015. We made 4 liquid cultures from each type of colony. We also made two liquid cultures from the glycerol stock of cells containing the ribosome binding sites B0030 and B0032. I plan to come in tomorrow to miniprep each of these 10 liquid cultures for eventual sequencing (in the case of the SAMT.B0015 and BSMT.B0015 cultures) and ligation (for each culture).