Saturday, November 22, 2008

On to Step 2

One of the things that I'd like to accomplish at work is to develop a certain kind of DNA marker, called microsatellites. I'm lucky that my boss is supportive of doing this kind of thing, and if it works with the mosquito species of interest, I could conceivably repeat the process for other species.

I found a really clear and straightforward protocol, and over the course of three or four rounds or ordering, I've gotten all the supplies.

Great, all ready to go. Usually, you start with DNA from one individual, but I quickly learned that one little skeeter doesn't yield enough DNA. So off I go, trying to get the right amount of DNA from several mosquitoes processed together. And right off the bat, I run into problems...

The first picture is from my early attempts. What you want is for the DNA to look like the middle bands of the ladder (shown as L, ignore L2). Nice and crisp, not smeary. Notice how the only one that sort of fills the bill is 6, and, unfortunately, it was another species, thrown in for comparison.
So I try this and that, and finally, find another protocol that has me process 10 bugs in an impossibly small amount of liquid, and wouldn't you know, it works. It works!! See the picture below, both lanes labeled D, and how they are actually brighter than the ladder (L), which means that there is plenty of DNA in the sample. No smearing shows that the DNA is of good quality as well.
Yippee!!! Now I can move on to the next step. Stay tuned.

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Hi, sorry to make the humans do an extra step.