Fiona

We arrived on Monday and had a tour of the vet college at NCSU,(which looks amazing!) before settling into our accommodation. The next day we went into our labs and found out more about the projects we will be doing while we are here.

My project is helping to look at how a particular gene is involved in the immune function of zebra fish.  This involves using Real Time PCR and gel electrophoresis to investigate the levels of expression of the gene. 

On Tuesday I was shown how to carry out the Real Time PCR and gel electrophoresis, I also learned about the importance of pipetting the samples accurately and ensuring that there is no contamination in any of the reactions to be certain the results are reliable.

Wednesday was Independence Day so we had the day off to explore Raleighand visit the celebrations downtown. Unfortunately we got caught in a thunderstorm, but we managed not to get too wet and it was dry in time for the firework display in the evening! I spent the rest of the week performing more Real Time PCR and gel electrophoresis to acquire more practice before using these techniques to start on the actual project next week.

On Saturday we visited one of the local shopping malls, before going out to a restaurant in the evening. Sunday was incredibly hot, with temperatures getting up to 41°c, so just right for a lazy day by the pool near where I’m staying!

Lucy

We departed from Heathrow on Monday and arrived inRaleigh in the afternoon. We were picked up and taken to the NSCU vet school where we were given a short tour and of theVetSchool and introduced to the people we were staying with and the people in or labs who we’d be working with. We then went off with our respective housemates and did some grocery shopping before going home to bed.

Tuesday was the first day in the lab: First I was given a talk on what the project I was going to be working on; what it was about and what I’d be doing for it in the next 9 weeks. Then we went into the labs to do some PCR on some genes (V, W, X, Y and Z) from the organism in our project. After the PCR I learnt how to prepare, perform and image an electrophoresis gel in order to check that we’d successfully amplified the genes using the PCR technique. Then I was shown how to do homologous transformation to make clones of the genes we’d amplified. These were plated onto selective media and then placed in an incubator and left to grow overnight.

After the lab: Fiona and I went out with Katie (the person Fiona is staying with) for a Mexican meal which was really good.

We had a whole day off work on Wednesday because it was Independence Day! So after a nice lie in, Fiona and I met up and Liz (the person who I’m staying with) dropped us into downtown Raleigh in the afternoon for all the festivities. The streets were packed with stalls selling food and stuff, there was wrestling, melon seed spitting competitions, eating competitions and much more.

We wanted to stay for the firework at half nine though so we walked through the stalls again up to the firework end of the street. When we got to the end of the street there was a circus act performing so we watched that, then there were bands playing which we listened to then the circus act came on again with their full show just before the fireworks. We then watched the fireworks, which were pretty good.  

After the fireworks the streets were completely packed with everyone trying to get back up the street and out so we were slowly making our way up the street and we happened to walk up to and then stop by an American film crew so we ended up on TV! Not bad after just three days in the country!

In the lab on Thursday: We checked our selective plates from the homologous recombination for growth. Three (X, Y and Z) out of our five had colonies on them. We reran the PCR and gel electrophoresis for the two (v and W) that didn’t work, but unfortunately it failed again (we could tell by the gel electrophoresis this time). So then we took a few of the colonies from each of the plates that did work (X, Y and Z) and cultured them in liquid media for using in restriction digestion (the next day).

Friday in the lab: We took the liquid media cultures that had been growing overnight and pelleted the cells and then performed mini-preps to extract the plasmid DNA.  We then performed restriction digests using endonuclease restriction enzymes to cleave the inserts and see if it was the gene of interest that had indeed been  cloned.

After the restriction digestion we ran a gel electrophoresis to determine if we’d had the right insert. One of them (X) had the right insert, but the other two (Y and Z) didn’t. At this point Henry (the person working with and helping me in the lab) had to go home so I was left to pellet and mini prep some re runs for the two (Y and Z) failed tests completely by myself and other than I slight malfunction with a pipette it all worked!

On Saturday I had another nice lie in then met up with Fiona and we went to Cary Town Mall for a bit off shopping. We looked around some of the bigger shopping centres then we all went out to dinner inChapel Hill at a place called The Restaurant on the Hill because Liz’s sister had come to visit for the weekend. We both had Mac and cheese withNorth Carolina slow cooked pork which was really good and the first time I’d ever had Mac and cheese!

On Sunday I got up early this morning to go with Liz to the barn to see her horse and help her out with the feeding etc. After we’d got back I went over to where Fiona was staying because there’s a pool there so we spent the afternoon swimming in and by the pool which was lovely especially as the temperature reached 106 ᵒF (41 ᵒC). Liz picked me up from there and her sister was still here so we all went to Fresh Berry which is a frozen yoghurt place. It was really good and just what you needed on a hot day.

 

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