It has been a difficult past few weeks in lab. Not only have I been the sole person in charge of doing most things (ordering supplies, maintaining the lab reagents and "cleanliness", taking care of our animals), I have my own project to think of. I would be fine and dandy with doing everything if only my experiments worked. I mean, having to do everything myself in lab is a very good experience to really learn and know how to run a lab in the future if I do go down that path. But it's especially irritating if my experiments didn't work! Then I start questioning my techniques, whether I was distracted/overworked or if I'm just plain stupid.
I've been working on RNAi viruses for more than a year and it has always showed some potential. RNAi (RNa interference) is basically a technique that many labs use to knock-down (or eliminate) very specific proteins. Thus far, when I've tested these viruses on liver cells, they've always worked. But now, when I tested them on fat cells (the kind that my lab is interested in), it isn't showing 100% positive results! Argh!
I'm currently repeating the damn experiment, hoping that I wasn't careful enough to measure and use the same amount of proteins, but it's highly unlikely (thought still a possibility). The second possibility is that our infection procedure/protocol isn't very suitable for fat cells. If true, I would have to probably read up and devise alternative methods or infect our cells with control viruses to see if it is infection protocol that is problematic. The third is that my stupid sequence is just failing to work well on my target proteins, which is the possibility that I'm dreading most! If the third is true, then I'd have to essentially start all over again and this will delay my graduation. Argh!
Sigh~ Wish me luck on the repeat experiment. Here's to hoping that I just made a minor mistake the first time round...