First bit of real research! How to make a LilBUBome – STEP 2. Sequence a target region

Lab Note #7
May 02, 2015

Hey All,

as we wrote you on Thursday, we've decided to do a lucky-guess experiment to look for mutations in a tiny part of LilBUB's genome, a region called the ZRS. So, this week we've been working pretty hard to get a sample ready for sequencing, because May 1st is a bank holiday here in Germany. Well, it turns out I was too slow so now we'll only get the result of what the exact DNA sequence of LilBUB's 800bp ZRS is on Monday.


So, what did we do?

First, we extracted the DNA from some of the blood sample. We then used a technique called PCR to purify (and amplify) only the DNA contains the ZRS, so that we have a lot of material to analyze (here's how it works).

After that, the first thing we checked is whether amplifying LilBUB's DNA worked just like amplifying any other cat's DNA (which I did as control in parallel - thanks to my neighbor Nadja for the sample). Turns out, BUB's ZRS (remember, the part of the DNA that controls the number of digits) is the same size as that of the control. OK, nothing unusual so far.


The amplified ZRS of a control cat (left) and LilBUB (right)


But what about the actual DNA sequence? We sent only only the amplified bit of DNA in the picture above for sequencing (0.00023% of BUB's genome) and until Monday we have to wait for the results. So let's see, my guess is it's like nothing unnormal in the sequence here as well.


Best

d



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