PROGRESS 20: planned out a new dry lab strategy for in silico mutagenesis! our time in the wet lab will be unfortunately limited, so we are finding alternate ways to get results and pushing ourselves onward.. #idec #directedevolution #syntheticbiology #synbio #imperialcollege

PROGRESS 21: tried out a few new bioinformatics tools today (80% of the process is setting up the software to work on your computer!), the tools seem promising to generate and analyse our mutants in silico #idec #directedevolution #synbio #syntheticbiology #bioinformatics

PROGRESS 22: tested Imperial’s bioinformatics server, ordered DNA Pol kits + primers, organised temporary lab space and equipment access for when Hackspace closes next week, then made more delicious&nutritious broth for our cells😋#idec #directedevolution #bioinformatics #synbio

PROGRESS 23: weekly team meeting discussing wet lab+dry lab+wiki progress, planned out goals & experiments for every day next week, inoculated cells containing plasmids to purify tomorrow, and consumed a bag of drumstick squashies #idec #directedevolution #syntheticbiology

PROGRESS 24: did some electroporation and microplate reading (the first time some of us have ever done these protocols and used these machines!). many of us haven't had the chance to do much wet lab work in uni because of COVID, and this has been a great opportunity #idec

PROGRESS 25: our primers are here! we also set up our workspace in the Chain Lab on campus where we will be for two weeks. #idec #directedevolution #syntheticbiology #imperialcollege

PROGRESS 26: we completed our first cycle of sequential PCR mutagenesis! Our protocol didnt exactly work.. BUT we managed to troubleshoot by running gels throughout the process and we’ve thought of solutions around this to try on Tuesday #idec #directedevolution #molecularbiology

Some early results we had from a dry run of our screening protocol on the well characterised LuxR: this is LuxR screened at an increasing concentration of AHL, the higher the concentration the more LuxR that can initiate expression of RFP

PROGRESS 27: designed primers for gene assembly, made figures for our report/wiki, analysed data from AHL screening, discussed higher-throughput screening methods, and celebrated team member Dana's birthday with cupcakes! 🧁 #idec #directedevolution #syntheticbiology #synbio

PROGRESS 28: designed another set of primers for an alternate method of gene assembly, designed a few pages of our wiki (but haven't coded them yet), and looked further into flow cytometry as a method for screening #idec #directedevolution #syntheticbiology

PROGRESS 29: had our weekly team meeting with catchups from wiki/drylab/wet lab, ran a pcr which failed so ran it again under new conditions, ordered new primers for an alternative method of cloning #idec #directedevolution #biology #molecularbiology

PROGRESS 30: performed Gibson assembly, transformed cells with un-desalted and desalted Gibson products, then plated everything (and made some bacteria art using excess agar and cells! 🦠🎨🖌) #idec #directedevolution #syntheticbiology

We plated 12 LuxR variants (with random mutations around the binding pocket) onto agar with different AHLs! Each colony with a different variant responds differently to the same AHLs. We can compare this to wild-type LuxR behaviour to see the effect of mutations #idec #synbio

PROGRESS 31: we used the Imperial Science flow cytometry facility to screen and sort for variants that can bind to a non-cognate AHL as well as the wild-type can bind to the cognate AHL! (photo: our teammate Lucy) #idec #directedevolution

We also said goodbye today to our team member Laima who will be going home for 2 weeks to visit family ❤️ she’s worked very hard for the wet lab team and we’ll miss her so much! She’ll keep contributing remotely for the rest of the month

We said goodbye to another team member this week! Aarushi will be heading home for the remaining days of summer. She’s been a great member of the wet lab team👩🏽‍🔬(we forgot to take a lab photo for her, so we drew her a lab on a whiteboard instead)

A belated congrats to @Evonetix for their recent patent on DNA storage and retrieval!! evonetix.com/evonetix-grant…

This paper by Gu et al (2020) is a great example of how the unique dual function of the EsaR quorum sensing transcription factor can be exploited for synthetic biology #synbio #syntheticbiology pubs.acs.org/doi/10.1021/ac…

After months of working together semi-remotely, the FULL team finally had the chance to meet in person yesterday. Thank you to @iDEC2021 for bringing together this team of amazing individuals!

🏆Very very excited to announce that the Imperial team just won the @iDEC2021 INDUSTRY ADVISORY GROUP AWARD for "the project with the most industrial application value"!! Special thanks evonetix, Genewiz Azenta Life Sciences, Imperial College Advanced Hackspace #synbio #idec #syntheticbiology