Today as I was walking to Dr. Cryan's office, Dr. Cryan was coming towards me and in passing said that he was just called into a meeting, so he couldn't work with me today. He brought me into the lab and introduced me to Sam and Julie. Sam was talking about computer software when I came in. They explained some things about how whenever a genome for a species is completed, it gets entered into this world wide data base that anyone can view. Then, Sam went off to do his work, and Julie worked with me for the day. We ended up making two gels. The first time, I did the same things that I've been doing, making gels, inserting the DNA with a pipette, and then running gel electrophoresis. Unfortunately, nothing showed up. There was however some thing shown in the picture. Julie said it was probably excess primers that were added in the PCR. Just in case, she said we should rerun the gels and see if the same thing happens. After making a second gel, we saw that nothing appeared. This means that yet again, something was wrong in the mix. After the second gel was done, Dr. Cryan came back from his meeting. He said that next time, since we wouldn't have time today, we should use someone else's kit, someone else's station and try it again. I guess a different person tried to run it and got good results. Then we went to his instructions and compared them to mine. He had added more magnesium chloride and doubled the amount of primers. Both Julie and Dr. Cryan said that they were important changes and could very likely be the reason that my gels haven't been working properly. Next week I will start all over from using the PCR with the other person's instructions, kit, and lab station.
It was nice working with Julie today. I was able to compare different techniques and processes that the two scientists do to accomplish the same tasks. For example, Julie uses a "cross linker" to clean some of the trays before we use them. It looks like a microwave. You just place the tray into the machine, and it uses UV light to clean potential excess DNA off the tray. Dr. Cryan just uses water to clean the tray out.
All in all, I am crossing my fingers that next week's test works because we are running out of ideas. I am trying not to be discouraged, but I guess we'll have to wait and see!
Nice post, Kara - full of detail and explanation. I am glad that you were able to do work even when your mentor was called away.
ReplyDeleteDid you know that Josephine works with Julie and Sam?
Please change the format of your blog, as I wrote about last week. See me if you have questions.