My relationship with phage lab is one of much uncertainty. One thing I am certain of is that it is testing my usual calm and positive attitude towards life. Science is very much like that and if I want to continue my work in the field of science I shall just try and get used to it. It can be frustrating and tedious and challenging. Phages and success in phage lab seems to avoid me at me at all cost. If you have read my last blog, you will know that I haven’t had much luck with phage lab but after I had gathered a lysate from my phage I hoped my luck would change.
DNA extraction is the next step of phage lab and possibly the most tedious, closely rivaling direct isolation. DNA is a molecule that carries our genetic code and in order to find out more about the phage I had finally we had to extract the phage’s DNA. DNA provides all of the information you could ever want from an organism. A protocol similar to what we followed can be found here. 
Luck continued to evade me and for some unexplained reason I could not find my phages DNA to save my life. I had to repeat the DNA extraction protocol 6 times in order to find enough DNA to move on to the next step. The first reading I got was so low that there wasn’t even any point in keeping the sample. The process takes around an hour of continuous work and so it wasn’t the easiest protocol to follow but by the end of this stage I had all but memorised it. The next time I got another low reading and I was thinking ‘oh no, here we go again’. I thought I had gotten passed the unluckiness that plagued me but alas it had reared its ugly head again. I ended up having to spin my lysate (the liquid that contained my bacteriophages) so that the phages concentrated at the bottom in order to get a greater concentration of the bacteria. Thankfully the next time I did DNA extraction I got a much larger concentration of DNA but I still had to end up doing it three more times.
I had gotten to the stage where I had almost had enough DNA to take to the next step and then suddenly a whole tube of DNA appeared one day in lab. I had labelled the tube as being DNA but I had no record of it in my log book nor did I know where the heck it had come from. If this DNA sample was actually DNA then I would have more than enough DNA to take to the next step and wouldn’t have to go through another round of extraction which had become mind-numbing at this point. I ran the DNA on a gel which would tell me whether or not I had DNA. An in-depth explanation of this step can be found here. Somehow my sample that had come out of nowhere was DNA and good DNA at that.
This is “Mooo”.
We got to see our phages through an Electron Microscope which was awesome as its really hard to see any progress when working in lab because you can’t see your phage and so to see it in real life was rewarding. The Electron Microscope we used cost over half a million dollars and so I was almost too scared to touch it, knowing how clumsy I am.
I tend to be a person who doesn’t follow the recipe book exactly while cooking and so there may be too much of one thing and not enough of the over. This doesn’t really matter when it comes to cooking but when it comes to science it is a bit of a problem. Following a protocol exactly how it is lined out in the book is really important because accuracy is key to success in science. I struggled with this precision which lead to many mishaps and mistakes made. It would seem that maybe science wouldn’t be the thing for me due to my clumsy nature and inability to precisely follow a protocol. Things are not always as they seem and I’m here to show you that even clumsy people can eventually make fabulous scientists.
The next step of phage lab now is to sequence the DNA of our bacteriophage. I have named my phage “Mooo” which is fitting as its appearance is short and stubby, kind of like its founder! Science requires one to be persistent and to just keep going and that is exactly what we had to do. It challenges you in a way that can make you question your sanity and even though it may not seem like it, everything will turn out okay in the end. It’s a fickle business but getting through the lab work and being able to see what we achieved in the end makes it all worth it.
Things can only improve from here, surely.
See y’all in the next instalment of this scientific adventure.
- Gill, J.J., Gill: Phage Genomic DNA extraction 2015.
- Addgene, Agarose Gel Electrophoresis.