Over the course of this summer, I invested a full month of research into attempting to verify the identity of the reporter plasmid – and I failed to do so. In fact, I showed rather conclusively that the plasmid was not, in fact, the backbone that we had been led to believe it was. Interestingly, we also learned several other things about the plasmid, including that it does confer Kanamycin resistance upon E. coli, that it contains at least one cut site for a restriction enzyme that was anticipated in the original plasmid, and that it has a length probably several times longer than what was anticipated.
From this knowledge, we have multiple options. Because we like to verify others’ results, our lab will reach out to the lab which gave us this plasmid originally and re-request that sample, so that we can replicate their usage of it as a tool. In parallel, we are designing our own tool for the same task – hoping to use standardized synthetic parts which will ensure that whatever we do is easily repeatable. Additionally, we have ordered and successfully cultured a third plasmid which has been used to serve a similar function, and are currently using that to test a transformation protocol. While this has been a setback, it has not stalled our research, and it has taught me a valuable lesson about checking my work at every step, and having the ability to verify the identity of a molecular product. Despite not having great success during this month of research, I enjoyed my work and am continuing to research in the same lab during this academic year, and I hope to continue researching in the future!