Keep track with what we did so far

(23.10.09)

This week we have done the following :

• Continue the characterization assay, identify the light induction time
• Using the results we got in the computational part of the project (see Modeling part), we mutated our LovTAP at ILE427PHE and LEU453GLY: we then took the ILE427PHE for further experiment to see if the time response to light activation was shorter

(16.10.09)

This week we have done the following :

• Clone LOVTAP into iGEM plasmid
• Parts registered & sent to iGEM HQ

(09.10.09)

This week we have done the following :

• We did a characterization assay with different time of exposure to the light
• We did a characterization assay with 2 hours of exposure to light, with the plate reader and the qPCR machine
• Digestion assay with the potential double transformant in the Trp K.O. strains

(02.10.09)

This week the following things were done :

• We tried to do a characterization assay of the entire system, but the LED were placed to close to the cells so it killed the cells
• We send all our parts for sequencing

(25.09.09)

This week we have done the following :

• Culture of Trp K.O. strains
• Transformation of Trp K.O. strains
• Tried to do some characterization assays, but it didn't work well

(18.09.09)

This week we have done less wet lab, as the academical year has begun (so we had to go to our courses):

• Further characterization of RO1 and RO2.
• Culturing of Trp K.O. strains

(11.09.09)

This tenth week of wetlab we have done the following things:

• RO1 has been characterized

(04.09.09)

This ninth week of wetlab we have done the following things:

• RO2 has been characterized

(29.08.09)

This eighth week of wetlab we have done the following things:

• Cultures to clone the biobrick in front of RO2 -> didn't work
• Double transformation
• Send to sequencing
• RO1 is yet not working

(22.08.09)

This seventh week of wetlab we have done the following things:

• Beginning of the RbphP project (PCR colony)
• Try of 1.5 step PCR to get the complete biobrick from LovTap -> didn't work
• Check (digestion assay) to confirm we have the LovTap-term
• Try of medium M9 for RO2
• Characterisation (fluorescence in fonction of induction) for RO2
• From LovTap-term, ligation to obtain the biobrick LacI-Rbs-LovTap-term with cloning steps (not from the 1.5 step PCR)

(15.08.09)

This sixth week of wetlab we have done the following things:

• Protocol of ligation was refined.
• !!! Inducible LOVTAP-term and Read out 2 biobrick were created.!!! The system is completed.
• They need to be sequenced, characterized and submitted to the registery.

(08.08.09)

This fifth week of wetlab we have done the following things:

• Problem in ligations.

(01.08.09)

This fourth week of wetlab we have done the following things:

• Our gene of interest (photoreceptor LOVTAP) was cloned in front of a terminator (iGEM part [http://partsregistry.org/Part:BBa_B0015 BBa_B0015]). We created our first biobrick.
• The second biobrick was coloned as well, but we wait for the results to confirm that the plasmid contain the right insert
• Modeling part: Preliminar simulations were lauched, and their analysis were done. Simulations will be lauched in few days.

(24.07.09)

This third week of wetlab we have done the following things:

• Again : Several attempts to create the first biobrick (Our gene of interest (photoreceptor LOVTAP) in a iGEM plasmid containing a terminator) and the second biobrick (A lacI promoter in front of an RBS), but no results were obtained for the first biobrick neither for the second one.
• Modeling part: Preliminar simulation was launched over the weekend.

(17.07.09)

This second week of wetlab we have done the following things:

• Several attempts to create the first biobrick (Our gene of interest (photoreceptor LOVTAP) in a iGEM plasmid containing a terminator) and the second biobrick (A lacI promoter in front of an RBS), but no results were obtained for the first biobrick neither for the second one.
• Modeling part: Files required to launch simulations were created and analyzed.

(12.07.09)

This first week of wetlab we have done the following things:

• Transformed the plasmids with the LovTAP gene, generously sent by Dr. Sosnick's lab from Chicago universtiy, into competent E.Coli
• Designed the cloning strategy for cloning the LovTAP gene from its original vector to a iGEM vector+add an inducible promoter (LacI) (+RBS +term.)
• Ordered and received the primers needed for the PCR of LovTAP
• Designed the cloning strategy for inclusion of the LovTAP BioBrick with different reporting cassettes
• Transformed all the BioBricks that will be needed for the cloning strategies (c.f. notebook for more information about these parts) into competent E.Coli
• Fused the two BioBricks "LacI" and "RBS"
• Digested the LovTAP PCR products and RBS part