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Dr Madhavan Jagadeesan

Date: 14th July 2018

Place: : Chennai

Dr Madhavan Jagadeesan is an ocular geneticist, molecular biologist & genetic counsellor. He additionally serves as the director of Dualhelix genetic diagnostics Pvt Ltd, Director of Academics & Research at Vasan Medical Research Foundation, and an adjunct Professor in the dept. of Genetic Engineering at SRM University, Katangulatur.

We had the honour and privilege of having a productive session with him on the 14th of July to discuss how we could improve our project this year. After the preliminary project explanations were given to him by our team members, he had two factors for us to consider. Firstly, our project was focussed towards screening the successful occurrence of substitution mutations. We had failed to consider the possibility of how our system would screen addition and deletion mutations and picked his brains on how we could achieve the same. We were able to use the suggestions we received and due to his timely help, we were able to make the necessary modifications.

Another query that he had with respect to our project was how our system would work when mutations were present in close proximity to one another and also when we wanted to screen whether larger genes of interest had been mutated or not. His inputs made us reconsider a few things with respect to our project and gave us a new perspective of understanding.

We plan on incorporating as many of his suggestions as possible in our working experiments this year. We hope that certain questions that we may not have been able to address get addressed by the future teams in the coming years.

Mr Phaneeshwar Rao

Date: 14th August 2018

Place: Chennai

On the 14th of August 2018, two members of our team had a very productive discussion with Mr Phaneeshwar Rao of Evolva Biotech about the project. Mr Rao is a molecular biologist working on mutagenesis and specialises in inducing multiple mutations.

We described to him the methods widely adapted for mutagenesis, namely PCR based and oligonucleotide based. As we discussed the differences between the two approaches, he also explained to us the fundamental uses of each in industry. We also spoke about the pALTER system which is widely practised today and introduced to him our idea of mutagenesis. We explained GFP, DFP and the mechanism of screening that FluroScreen would employ. We showed him a few results that we had obtained during InterLab transformations and asked him for comments and on our project.

The suggestions that he gave our team were twofold, the first being that we add two primers, one for the sense strand and the other for the antisense strand of a double stranded plasmid. The second input was about how we could compare the efficiency of primers binding to a single stranded plasmid and double stranded plasmid and in fact study it as a part of our modelling objectives. This input arose from the doubt on the stability of single stranded plasmid. Another valid consideration he gave us was about the decreased gene expression levels upon increased number of mutations to one sequence. He offered to help us with primer design and PCR optimisation and was willing to troubleshoot aspects of transformation and mutagenesis if such a situation did arise.

We are truly grateful for his suggestions and thank him for his time. We also plan on incorporating his suggestions into the working model of our project.