18
Oct
2011

My last week in the labs has been a fairly quiet week compared to some of the others. I have still been busy doing PCR and gel electrophoresis and now feel confident in both of these techniques. This will help me greatly in my next year at University as well as in my future career.  I have been carrying out PCR and gels on a variety of sample types including DNA from tissues, DNA from blood and DNA from serum from a variety of hosts including humans, cats and dogs.

I will be sad to leave the labs as it has been a really good experience on the whole and I have learnt so much during my time here, but I am ready to go home and see my family and friends again. This weekend we are heading up to the mountains for a final flourish before we leave.

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18
Oct
2011

Again this week I have been continuing with PCR and gels for various studies.

I have been testing samples from the study previously described looking for the competency of the cat flea as a vector. In this study Bartonella henselae is used as the infectious agent and to see if the fleas have passed it from one animal to the next. This means that all of the positive results we get should be B.henselae positive. However, for a few of the samples I have tested I received positives for Bartonella koehlerae, they also can back from sequencing as B. koehlerae. This means that somewhere during the experiment contamination with B. koehlerae has occurred. If this is the case it means that all of the experimental data will have to be scraped and it may have to start again. They will have to work out how B.koehlerae has entered the experiment and on first thoughts it is believed that it may be due to the fleas which could have been infected with B. koehlerae before the experiment started.  

Our adventures

This week we did a Segway tour of Raleigh as we thought it would be the perfect way to see the sights of the city and downtown area. We took an hour tour which took us down the main street, to the governors’ mansion and to other important areas of the city. On the same day, Bugfest was going on in the city. This is a celebration of bugs and there is even a bug cafe which serves chocolate coated crickets and mealworms in sauce. Me and Jade did not try any of the things as it didn’t look like the most pleasant thing in the world. However, we did get a funnel cake, basically a giant sugary doughnut!! Sparkfest was also going on in downtown and this is a celebration of the arts, people were doing chalk drawings on the street and there was various music and dancing happening down the main street it was a really good atmosphere although trying to dodge all the people and artwork whilst on a Segway was difficult to say the least.

On Sunday we decided to go to the cheesecake factory for one last time before we left. We decided that we would just have cheesecake this time as in previous occasions we have had other food and been too full for cheesecake even if we have only shared a starter!! We even got a piece to take home for the next day so we could try as many flavours as possible!!

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18
Oct
2011

 

This week in the labs I have been continuing to test various samples using PCR and gel electrophoresis.  This includes samples from the study I have previously mentioned, where I was testing for competency of the cat flea as a vector for Bartonella Henselae. This week I have also carried out manual DNA extractions on a few pieces of tissue. This process involved lysis of the tissue using protinase K, then cycles of adding various buffers and centrifugation. One of the samples yielded a very high DNA concentration as measured by the nanodrop spectrophotometer, so I had to repeat the last step in the process in order to dilute the DNA concentration as otherwise it would be too high to complete PCR and Gel electrophoresis and the results gained would not have been accurate.

This week I have also received the DNA sequencing results back from the positive samples I had obtained from the Canadian dog blood. Unfortunately, all the sequencing reactions failed to yield sequences that matched what we were looking for. This is a shame as it would have been nice to have been got one positive from all of my samples tested. However, this could be a good thing in the long run as this may mean that Bartonella species, especially Bartonella vinsonni subspecies berkhoffii type 4 has been eliminated from this area. However, more information is required before this assumption can be substantiated, including repeating the DNA sequencing.

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