Tuesday 30 September 2014

D-N-yay!

It's taken a few headaches, but I have managed to get COI (cytochrome oxidase 1, a mitochondrial gene/section of DNA) to work so I can get sequences. There were a few hiccups at first - the negative (control sample that I run with water rather than spider DNA, to test for contamination) showed contamination, and the primers stuck to each other and themselves and basically anything that wasn't the DNA, and I used the wrong combination of primers for a bit.

So now I can show you the process from beginning to end, in infographic form.


My breakthrough with COI is not only encouraging and positive for my work (now I just have to get EF1g working and ITS working more reliably) - it's entrancing. I have collected spiders, and I now have sequences from them. I feel I have grown the wheat, milled the flour and baked a delicious loaf. I can't really get my head around how each wee spider is made up of heaps of little cells that contain sooooooooooo much information. Before now, I had collected samples, I had extracted DNA from samples, and I had sequenced DNA - but never all on the same sample. For some reason a huge part of me just assumed someone was doing some magic somewhere along the process that made it work, but this time I've made it work, and there was no magic involved - just lots of baking.

That really is all that one has to do to obtain DNA sequences.

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