My Beginning in a Nutshell

         Hello, my name is Kristine and I'm currently enrolled in State College of Florida's Biotechnology program. I have always been in love with science and remember watching documentaries and television shows wanting to work in the labs that the actors/featured scientist were in. I thought in order to get to work in such a place you needed to go to Harvard or some high class university first.

            When I started my journey back to school I was working as a certified nursing assistant (CNA), and majoring in nursing. CNA's are the low man on the health care totem pole. In the beginning it's what gave me the drive I needed to get my nursing degree. My anatomy and physiology professor would tell us crazy stories of her lab days as a grad student and that sparked my childhood love for science. For two semesters I would hear these stories and wish there was a program I could take to learn these techniques. Working in a lab sounded so much better then health care to me.

            I was one semester away from completing my prerequisites for the nursing program when I caught wind of SCF's new Biotechnology program. After realizing that most of the prerequisites I had already completed satisfied the requirements for this program, it didn't take much to convince me to change majors.

            Now getting ready to graduate from this program I am both excited for the career and future ahead. As well as nervous as I'm being “kicked out of the nest” so to speak. I love being in the lab and I’m fascinated on a daily basis by this microscopic and molecular world that people tend to take for granted. This blog is a compilation of what I've learned, learning, and my “oops” along the way.

 

RSV and NIH-3T3

            The start of my fall term, I was in a plant and animal tissue culture class. In which we adopted our own cells to take care of for the duration of the class. I chose to work with NIH- 3T3 fibroblasts.

      

 

 

They were growing so beautifully! I passaged them 1:3 and to my surprise when I came to check on them the next class, I could count a total of 10 cells in the entire flask. Immeditatly I'm going through my procedure (thank God for notebooks!) just to makes sure I didn't mess up while splitting my cells. I finally called my professor over to take a look.  Of course I was a little on the nervous side thinking, Great I just totally failed this class. She was even confused as to what might of happened to cause the cells to be so few. We ran test to see if our media or trypsin could have been contaminated. But they came back negative. As I monitored the cells over a few days they slowly began to grow, but they looked very sickly. They also began to form syncytia. As nervous as I was about not getting a good grade, I was (in a mad scientest sort of way), excited that something weird was going on with my cells. My professor suggested they might have some sort of virus. That’s when everything seemed to make sence. The day before my daughter had been diagnosed with respiratory syncytial virus and, being the loving daughter she is, she had shared her germs. When I had passaged my cells, I didn’t know that I was harboring these germs. Even though I had used sterile technique, we learned that RSV can stay on the surface of gloves for a long period of time. We performed an actin staining in these cells which showed the syncytia as well as granules that corresponded with granules seen under an inverted microscope.

                   We are still researching and running experiments to determine wither or not this could be RSV infecting these cells. I’m trying not to get my hopes up about the results, but I would be lying if I said I hope the results don't show an infection.

 

“Don’t forget the Gel Red

 

            One of my favorite “oops” stories from my course would have to come from what we call the RNA class. Officially called Biotechnology Methods 1, it’s a class focusing on the purifying, quantitating and isolating RNA. After a two week break it was so nice to be back in the lab. We began this term by running a mutagenic PCR experiment. For a quick analysis we set up  an agarose gel. This was the first gel of the term, and I was “in the groove”. My gel was poured and almost solidified when I overheard another one of my class mates confirming with my professor on how much gel red to use. I had completely forgot to add the gel red into my gel before it solidified. All I could do was laugh about it as I starting making another gel. For some time after, every time we poured an agarose gel, at least one of my classmates would come by my bench to make sure I added the gel red.

   

     

      

      

Welcome Kristine!   You make

Welcome Kristine!  

You make a good point about inadvertently infected human cells with a virus from home.  I hope your daughter's doing better than your cell culture.

 

I also have a question - what is gel red?  I used to do lots of agarose gels, but it sounds like this is a new twist.  Can you describe what it is and what it does?

Thanks!

Sandra

GelRed is a safer version of

GelRed is a safer version of Ethidium bromide for staining nucleic acids. We would put about 2-3ul of it once the agarose was dissolved into solution, before we poured it into the casting trey, this way it would contain the stain and make it easier to dispose of it. Since I forgot to add the stain, my samples wouldn't of showed up under UV light.

 

K.A.Cerbone

Interesting, thanks!

Interesting, thanks!

http://biotium.com/technology

Hi Kristine! It's great to

Hi Kristine!
It's great to have another student blogger. I feel your pain with the gels. Last week, for some odd reason, I decided to make mine with water instead of TBE. The results were not pretty.

Keep up the good work and have fun blogging!

Mandy

I've been surfing online more

I've been surfing online more than 4 hours today, yet I never found any interesting article like yours. It is pretty worth enough for me. In my view, if all webmasters and bloggers made good content as you did, the web will be much more useful than ever before.

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