OH - THIS WILL BE EASY...

Famous Last Words!!! Well actually, far from the last words I've used since embarking on a new experimental technique I found in a published methods paper. While investigating the role of my favorite gene, I have had some interesting observations that has led me into a new scientific arena. Mitochondria.

Ah, mitochondria, I love the buggers. Needless to say: I'm hooked. Unfortunately I (nor anyone else in my laboratory) has much experience with this field nor many of the nuances of working with a semi-autonomous organelle. Fortunately, I have recently come across a methods paper which nicely details a protocol for measuring the amount of mitochondrial DNA damage using "Long-qPCR". As I am a trained molecular biologist, PCR doesn't scare me a bit (ha-ha-ha), here's something I can do quickly and easily. Since embarking on this seemingly easy protocol a month has progressed. I have managed to produce one set of data that very nicely fits our hypothesis, never to have the experiment work again... This is difficult, postdocs are supposed to be data producing machines. Gone are the days of learning and floundering. It's time to produce results and publish papers. As much as I would love to explore and discover in the field of my new found pet (the buggers) - I must produce.

So, my solution? Spend Spend Spend. So far I've ordered six sets of primers, two different enzymes, and isolated more DNA than one can shake a stick at. Maybe, I will buy my way out of this problem! One of these variations must work! No luck yet. But tonight I've found something new, another published protocol! This one takes advantage of my favorite piece of molecular biology equipment:The ABI7500. Quantitative real time PCR. Now there's some data to analyze. This paper's got the primer sequence, the enzyme, the cycling conditions, and starting template concentrations all worked out. I'm ordering all the reagents first thing tomorrow morning. OH - THIS WILL BE EASY...