S4 39: Metro Tech Presentations

Metro Tech Presentations 
As we wrap up the spring semester, I am trying to get to a good stopping point in my research. It looks like I have about two more procedures to complete and then I will resume this project at the start of the fall semester here at ASU. I have also spent time putting the final touches on my research poster in preparation for the Metro Tech event. I am looking forward to seeing everyone's research next week and spending some time to catch up with everyone in the program. After the semester is over, I will be spending the summer doing an internship at ASU-Tempe.

S4 38: Next Experimental Steps

Next Experimental Steps
After a successful assay, the next steps in the experiment require re-streaking colonies onto other plates. After the colonies grow, then we will perform a mini-prep. The purpose of a mini-prep is to extract the DNA plasmid from yeast and E-coli cells. During this experimental step, we will be extracting the plasmids from yeast cells. I also think that this might be a good stopping point in the semester. I will resume this experiment this upcoming fall semester.

S4 37: I love Liquid Nitrogen

I love Liquid Nitrogen

I am getting to the point in my lab project where I get to do an assay again. I remember liking this part in the experiment because I was able to use liquid nitrogen. This procedure is a little longer than an alternative procedure that we found but I like using LN so much that it's worth the extra time in the lab. The point of this procedure is to allow us to identify if a colony of yeast cells express the β-galactosidase enzyme expression. When β-galactosidase bind to the X-Gal, it cleave the molecule which then makes a blue pigment, which is very evident. This activity indicated that there is the LacZ gene was transcribed and translated. 

S4 36: Back in the Swing

Back in the Swing
This week in the lab, I went through all 30 plates to determine which colonies were abundantly grown on our -T,L plates that had added 3AT and Bexarotene. I was able to pick off 54 colonies to use in our next phase of the experiment. The process I completed this week was streaking the 54 colonies onto -T,L plates to see which colonies will regrow. After that step, I will perform a Lac Z assay on the remaining colonies. I am a fan of the Lac Z assay because that means playing with...I mean "utilizing" liquid nitrogen during the experiment.

S4 35: Too Hyped to Science

Too Hyped to Science
It was a bit of a struggle to be in the lab today but it wasn't because I didn't want to be there. This entire week I have struggled a bit with maintaining my attention span and felt like a squirrel. I have also been pretty fatigued this week by all the school work. I think it showed in the lab today when I kept having to restart certain steps over and reread the procedure every 5 seconds. I even lost an hour of work by making a tiny mistake that I usually don't make. It was definitely my most spacey day in the lab but I got through it.

S4 34: Y2H Website

Y2H Website
Last week, I worked on starting the Yeast Two hybrid experiment by completing a bait transformation into yeast cells. I let the cells incubate and had a successful transformation. The next step will be to transform the prey plasmid with the bexarotene compound. However, this week I have spent my time working on the Yeast Two Hybrid website with my research mentor. The goal of the website is to have a process where students and other professors can access and learn about the Yeast Two Hybrid procedure. My favorite aspect of the website is the flow chart that we are working on so that people can have step by step guidance as they perform their own Yeast Two Hybrid screens.

S4 33: Next Phase of the Project

https://pubchem.ncbi.nlm.nih.gov/compound/Bexarotene

Next Phase
The next phase in the project includes beginning the Yeast 2 Hybrid experiment again using the same DNA prey library that we made last semester. However, this go around we are going to add a bexarotene rexinoid to the bait and see if it promotes the same type of protein binding or additional protein binding. I am looking forward to repeating the experiment because it will allow me to practice the Y2H processes again. I will also get to refine my research notes on the procedures which helps with the Y2H website that my research mentor and I are working on.