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.

S4 32: Sequencing Update

                                                                                   

Sequencing Update

This week in the lab, we got the results back from our plasmids that we sent to get genetically sequenced. They came back without any problems so I guess my dilutions were right. I am glad! It's a funny reminder of how I need to be more confident and trusting of myself in the lab setting. Out of the four samples we sent to be sequenced we got matches from the Blast data base. The next steps of our project are to repeat the yeast two hybrid with added concentrations of Bexarotene and see if we get the same type of RXR binding as we did we the current sample. But for now, I get to enjoy spring break first. 

S4 31: BRB

Be Right Back
This week, I didn't really see lab time because I have three midterms next week so I have been hiding away and studying. What I have focused on this week was gathering information on my Yeast 2 Hybrid project in preparation for the project backgrounds we have to write. This week has been pretty intense with the work load. I am trying to push through and keep telling myself that spring break is one week away. It didn't help that I thought last week was the last week before break but life goes on. I can't wait until these midterms are over and I can get back in the lab.

S4 30: Rough Day in the Lab

Rough Day in the Lab

Let's just say I am glad today is almost over. One of the things I hate the most is not remembering how to do something that I know I learned and should know how to do, especially if it is easy. It also made me question if I am cutout for research.
Basically, I needed to perform a few dilutions before I could get the right volume concentrations for my experiment. I started with my M1V1=M2V2 formula and got some tiny numbers that made me question the answers. After over an hour of trying to remember, getting frustrated, self-doubting, and venting about how much I don't think I'm cutout for research, I think got some maybe half correct calculations. I hope they are right. Today helped me realized that doing dilution calculations for chemistry homework doesn't really work the same in the lab.  I realized that sometimes you have to scale up, scale back down, and then scale up because the lab equipment doesn't measure smaller than 2uL on a micro-pipette.

S4 30: Trouble Shooting

Today in the lab, I focused on trying to do some trouble-shooting in order to extract the third and final plasmid to be sent for sequencing. My PI is thinking that there is possibly a mutation in this sample that is causing cells to produce its own LacZ and histindine. For this reason, we believe that this is why we were seeing blue cells when we performed the Xgal Assay awhile back. So, I am going to try two more transformation with different DNA concentrations to see if we get sufficient results this time. If not, we are going with the mutation idea and sending our other two samples for sequencing and calling it a day.

S4 29: Another Day in the Lab

Another Day in the Lab
I have to admit that one of my biggest struggles this past semester was keeping good research notes. I would typically find myself using my research notebook for mostly notes and it was challenging to keep everything in sections since we aren't suppose to skip pages. So last semester I thought by keeping notes and procedures on the google drive would solve my problem but it did not. I then began to struggle with having notes and procedures in multiple places and I found that the disorganization ultimately affected my productivity in the lab. When I sought guidance from my research mentor, she suggested a few resources I try out and supported my desire to have an electronic research notebook. I familiarized myself with Microsoft One-Note and I am so glad I did, because its design layout was exactly what I had in mind! I am now able to have the app on my phone too, and it really allows for me to be more time efficient in the lab and refer back to my notes easily. My favorite feature is being able to take a photo of lab procedures on my One-Note mobile app and also instantly automatically sync it to my computer. Then, I can quickly add notes directly onto my photo. I even started using one note for my classes this semester, and I definitely recommend everyone to try it out.

S4 28: Wow! How Time Flies!

https://www.clipartmax.com/download/m2i8i8G6i8H7N4A0_time-flying-cliparts-time-flies-clip-art/

Wow! How Time Flies!

I still can't believe that winter break is over, and it's almost been two weeks back into the spring semester. This semester I will be continuing my work with Dr. Marshall at ASU-West focusing on Yeast Two Hybrid. I believe that this semester we are going to be testing how different compound concentrations affect the different protein reactions in the Yeast Two Hybrid experiments. I could be a little off as I am still learning about this myself. I am looking forward to getting back in the lab. However, I had to go back to work full-time this semester and am a little nervous on how the semester will turn out. Wish me luck!