In the lab: We had our weekly lab meeting to decide what to do next with our projects. We decided that the vector we’d made was good to go on to use in homologous recombination experiments. However, at some point we would need to send it off for sequencing to check it is completely right. Megan had set up a homologous recombination of our gene V (that we’d got from our new serial dilution of primers for PCR method) and gene W (which was from the normal version of the PCR). The bacteria containing the construct with gene W hadn’t grown on the selective plates so it had not worked at all and only one colony was recovered for gene V. So we picked that one colony and grew it up in selective liquid media overnight ready for digesting the next day.


In the lab: our bacteria containing the construct with gene V grew in the liquid media.  However, when we did a restriction digest of it, it turned out that is wasn’t the desired clone so that had failed. So we needed to try homologous recombination with it again (and with gene W). But we decided to try and get a more successful amplification of gene W first by doing our new optimized primer dilution PCR with it. So we did that and pooled together the best PCR mixes to make our gene W gene stock.

Lunchtime: Got a free lunch today at the lab! There was a laboratory company selling their products and to entice people in they were giving out free NC BBQ.


In the lab: We retried the homologous recombination of gene V and gene W, but this time we used fresh competent cells and used two different ratios of insert to vector (one was the one we’d already been using (3 or 4:1)and the other was a simple 1:1 ratio) to see if we got better results. So they were all plated on selective agar overnight.

Lunchtime: We went out for lunch again today. This week we went to Mitch’s Tavern which is and American pub. It was buzzing in there and amazingly busy. I got the Mexican chilli which came with cheese, salsa, sour cream and tortilla chips along with the bowl of chilli itself.


In the lab: All of our plates had grown a fair few colonies on them! Big success! By looking at the numbers of colonies for the different ratios we’d used there didn’t seem to be any correlation there so we put our previous failures down to old competent cells. So we picked three colonies for each gene and grew them up in selective liquid media overnight.

After the lab: Fiona came back with me on the bus and we had our now weekly, Ben & Jerry’s then we went toRidgewoodshopping centre to look at some NCSU merchandise.


In the lab: All three of the colonies for each gene had grown up in the liquid media overnight. So we picked two of each of them to go on to pellet and restriction digestion. So we did the pelleting and restriction digestion. Our results showed that we’d successfully got a clone of gene V (at last!). We hadn’t got one of gene W.  However, we still had one last colony in liquid media to try first before concluding failure. So we took that last culture, pelleted it and did a restriction digestion. The results showed that we’d also successfully clones gene W as well! This was a great result as we can now move on with the project! So we made bacterial stocks of all the successful genes ready for Monday.


We went toCrabtreeValleymall again today. To start off the day we had a pretzel for breakfast. I’d never had a pretzel before and got a cinnamon sugar one. After our pretzels we went to Alumni Hall which was the shop we’d seen the other week which sold loads of NCSU stuff.

We wanted to go to The Cheesecake Factory again but didn’t think we could fit any other courses in other than the cheesecake itself. So we went to the take out cheesecake counter in at the side of the restaurant instead of eating in. It was incredibly hard to choose but in the end Fiona chose the Oreo flavour Cheesecake and I went for the Mango and Key Lime Cheesecake. So we got our cheesecake and went to sit outside in the sun to eat them. My one was very good; I’m not entirely sure what a “key lime” is but it tasted nice and citrusy and I thoroughly enjoyed my cheesecake, definitely going to try key lime things again.


We went toCameronVillageagain today to look around the little shops there and to get some things to eat. We were a bit hot after walking from the bus stop toCameronVillageso we decided to go to Goodberry’s to get some frozen custard first.

As with most places here inAmericathere was a huge amount of choice. In the end Fiona went for a chocolate brownie sundae and I went for the days special parfait which was vanilla custard with raspberries and blackberries layered into it, topped with cream and a cherry. Afterward we went to look around some of the little gift shop type places. First we went into Ten Thousand Villages which was a shop which sold fair trade things from all round the world. Lastly we went to Sugarland which sold cupcakes, ice cream, drinks and some other sweet pastries. 

There were loads of cupcakes, many unusual flavours as well so it was very difficult to choose. I went for crème brulee flavour one (which had a raspberry on top and in the centre) and a mango flavour one (sugar sweet on top and a bit of mango in the middle). Fiona got a lemon flavoured one and a strawberry cheesecake flavoured one. They were very messy to eat as they had a massive swirl of flavoured cream on top of them which got everywhere and fell off whilst you tried to eat it!

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