Isolating DNA - Much Harder than it Looks on TV

A lot has been done this week, though there isn't really much to show for it. Who knew DNA isolation was a trial-and-error process? The best thing that happened this week was getting to pluck my eyebrows for science. The worst was... well, I'll think of something.

We're following a rough protocol for DNA isolation from human hair and it took a lot of research to find something mildly promising. These "recipes," if you will, appear highly guarded; most instructions involve the purchasing of a kit which contains a mysterious patented variety of solutions. However, my mentor and myself managed to find some instructions for creating a buffer solution to dissolve the eyebrow hairs. There is also a protocol for the isolation of DNA using phenol-chloroform, but we haven't gotten to that step yet because we were out of phenol-chloroform and had to prepare our own.

Instructions:

Preparation of Buffer XI Solution

There are some tips I've discovered in my search for an appropriate method:

* If you don't have to use hair, don't. From the material I've read, it's extremely difficult to get usable DNA from hair samples. If you have to use it, use eyebrow hair and make sure it hurts when you pluck because then you're more likely to have the necessary skin tag at the bottom of the hair.

* Get a kit, or buy already prepared phenol-chloroform. It is a long, time-consuming process that involves highly toxic substances, namely, phenol and chloroform. My mentor and myself enjoy living on the edge, so we're making it ourselves, but we don't know if it has even worked yet because our first attempt failed and we haven't completed the DNA isolation yet; the phenol we had thawed was a pinkish/orange which means it is no good.

On another note, I'm sure many of you are curious about what exactly Epidermolysis Bullosa Simplex is. I will be explaining in my presentation tomorrow, but I can give a brief overview. In summary, it is a genetic disease characterized by fragility of the skin and blistering; severity varies among the four most common phenotypes, as well as between family members. It is usually inherited in an autosomal dominant pattern and is most commonly caused by mutations in the genes KRT14 and KRT5, both responsible for making different forms of keratin. This molecule is important for the functioning of the cytoskeleton. When it isn't working properly, the epidermis and the dermis separate, hence the blistering. There is a website below that I've found very helpful; there's even a section with information specific to genetic counselors. 

Epidermolysis Bullosa Simplex (EBS)

There are also some pretty interesting (if not gruesome) pictures of those afflicted. If you are in the least bit squeamish, I do not recommend clicking on the links. Some pictures are of babies, as some phenotypes manifest at birth. I'll be using these photos in a presentation tomorrow but they can also be found below:

Weber - Cockayne (mild)

Koebner (moderate)

Mottled Pigmentation (moderate/severe)

Dowling-Meara (severe)