Team:EPF-Lausanne/Last News

Last News

=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 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

