Team:BostonU/NotebookQS

From 2013.igem.org

Revision as of 18:39, 22 July 2013 by Pshah23 (Talk | contribs)



Quorum Sensing Notebook

June 2013

We planned out that we will transform E. coli with the civI/civR system found in Chromobacterium violaceum. We will design MoClo devices that include these systems. In the meantime, we are beginning to design primers that will make the civI/civR system into a MoClo system by adding the the fusion site in addition to the BBS and SpeI sites. Then, we will develop a PCR strategy.


June 18, 2013

Our advisor, Traci, emailed different labs and PIs that work with Chromobacterium violaceum. Dr. Jim Thoden from the Holden Lab at the University of Wisconsin sent us genomic DNA.


June 20, 2013

We designed forward and reverse primers for CviR/CviI systems and ordered those primers. We are currently waiting on acquiring the primers to begin PCR.


July 8, 2013

Today, we did Phusion PCR using the genomic DNA of Chomobacterium violaceum as the template for the CviI and CviR systems. We ran the PCR product on a gel and the CviI did not appear on the gel. The CviR system appeared and we gel extracted the parts.

The gel concentrations are as follows:


June 9, 2013

We redid the PCR for CviI. We ran the PCR product on a gel and we did the gel extraction. we quantified the concentration on the NanoDrop.

We ran a Level 0 MoClo reaction with the higher concentration of CviI and CviR.


June 10, 2013

We did a transformation using Bioline Gold E. coli and plated the samples on IPTG + Xgal.


June 11, 2013

We pulled the plates and picked three colonies and plated them on a stab plate and set up liquid cultures for incubation.


June 12, 2013

We set up CviI_CD and CviR_CD for sequencing in triplicate.


June 13, 2013

We analyzed the sequences for CviI and CviR and all of the samples we sent in matched the expected sequence.


June 17, 2013

We made glycerols and archived the highest concentration of each CviI and CviR.