S-STEM Scholar Marisa Ostos Blog

Since transformation of our Deinococcus radiodurans fell through, this week's focus was on finalizing the process to this time run three sets of transformations (including the creation of all broths we will need for inoculations, nanodrop readings, plating, and more). So far, we have tried running transformation three separate times (each time becoming a bit faster and a bit more precise in our methods), but since we didn't achieve transformation in each one, we decided to take a step back to try and see if we could try something different.  Our classmate had previously been able to run transformation successfully with a plasmid smaller. So to make sure that we actually are following the correct protocols, we will be running three transformations with three separate plasmids (and using Deinococcus radiodurans), one of which will be the smaller plasmid used by our classmate to perform his own transformation. If we are able to achieve transformation at all (especially if we can d...

  This was the week where after running transformation and plating on Friday (a third attempt), we were hoping that we would have transformed Deinococcus radiodurans such that it would have shown antibiotic resistance via integration of the pRADZ3 gene into its own genome. However, this unfortunately didn’t happen, for reasons that we are unsure of (so far). When we checked our plates on Day #4, 5, there was still no growth, indicating no transformation. Because of this, it is time to go back to the drawing board and are looking over every piece of the transformation protocol, from inoculation of E. coli (with its necessary plasmid that will be extracted for transformation) and   inoculation of D. rad (which is the species of Deinococcus that will be used for transformation) to plating of the hopefully transformed bacteria.   Now, an interesting factor to consider while looking at steps to transformation [on paper] for a third time   is to consider a factor that ...

  This week, our major focus was on ensuring we understood every part of our transformation protocol (especially in regard to our controls) so that we could ensure a [hopefully] successful transformation! Now, one important concept that our mentor Jonathan helped us to understand was the importance of proving the viability of our competent cells (taken from freeze back), which would help to show that our competent bacteria weren’t the problem if we had no growth after transformation, and proving the stability of our environment, which would help to show that the plates held a solid environment for D. rad to grow in. Because transformed cells need to be plated after 16 hours, and because an entire weekend had passed since we transformed our cells with the pRADZ1 and pRADZ3 plasmids (in separate groups of course), we would need to restart part of the transformation process (after ensuring that we fully understood each step in our transformation process of course).  We did this ...

Welcome to the first of many blogs that will provide you insight into the world of transformation (my current research endeavor), where the team I am on will be working on importing “superpowers” to an organism whose natural abilities would likely already put Superman’s to shame. Now, I’m not here to debate about which species would win in a match (Although doesn’t one by the name Deinococcus sound much more tenacious, sophisticated, and enigmatic than a Kryptonian species that is just so 20 th century)? But I am hoping that, by incorporating both illustrative and text-based pedagogy, you will join me in my journey of undergraduate research.  To provide a brief background, I joined Dr. James Tuohy’s Independent Research Lab at GCC just three months ago, and was excited to learn that I recently received the TRAIN Grant to help fund my educational expenses (thank you, Dr. Tuohy and Co!). Since then, I’ve learned quite a bit about what it means to truly put the Scientific Method to...