Preparing the DNA for sequencing, Shane Yeager

Interviewee: Shane Yeager. Shane Yeager, from the Whitehead Institute Center for Genome Research, explains the processes of storing and preparing DNA for sequencing. (DNAi Location: Genome > The project > Players > Public > Preparing the DNA for sequencing)

We take the DNA in, in E. coli out of our freezer and take that to our DNA preparation team. They'll go ahead and grow that big bacteria up so we have a higher amount of DNA to harvest eventually and then we'll break those cells open, get the DNA out, and the DNA will be broken apart by what we call a hydro-shear. It's very simple in theory, it's very similar in fact to trying to get a clog out of your toilet with a plunger. You go ahead and use shearing forces to break apart the DNA, so we have these random little bits of DNA to insert into our, our E. coli later on in the process. That goes on to our library construction team, they go ahead and put the DNA into the new E. coli cells by a process known as transformation. We link it all together so it's a nice solid little plasmid, and then it goes on to core sequencing. We grow it up a little bit, while continually shaking it so that it doesn't attach to the sides of our plates. And then we go over to the cubots, which automatically pick out individual colonies on a plate. From there we grow them again, again always trying to increase the amount of DNA that we're going to harvest, resuspend them with a little bit of water, purify them two times, once to get all the E. coli DNA away from the human DNA, and the second time to collect all the human DNA.

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  • ID: 15162
  • Source: DNALC.DNAi

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