After I had entered research phase of my degree I was always planning how would things go. I would rehearse everything in my head so that when the day finally arrives, everything falls right on place. The biggest hurdle was to get 1000 numbers of stress free, disease free Catla, my experimental fish. I was constantly calling people, fish farms to inquire if the fish was available but to no avail. Finally, one day a batch-mate called to say that he could find one Catla among his stock of fish (some other species). I was excited. I knew that I had to start with this at least.
The fish was brought from him to our lab, kept in an aquarium for acclimation. I had to begin with extraction of hepatic RNA. All I had was one fish and only one attempt. Every thing had to be righ else the things would fall back to naught again. While the fish was acclimatising to lab conditions I tried sacrificing other fish which was easily available and used its tissues to extract RNA. I found the process very simple and got good quality RNA. I was happy but one more shot was required.
When everybody was busy celebrating diwali, I was in Lab with fish tissues extracting RNA. I ran agarose gel electrophoresis. It was midnight by then and whole India was immersed in deafening noises of crackers and choked with sulphurous fumes. But I was in the silence of our lab with my agarose gel under Gel-Doc Imaging system. All I could hear was sound of CCD camera in the system trying to track gel. A blurr on the monitor and then with a flash I saw clear bands of ribosomal RNA. I was happy. I didn’t need crackers to express my happiness. I knew that next day was the real test where I would finally use my fish.
I woke up at 6. The earth was calm as everybody was asleep after a late night Diwali celebration. It was warm but smoky everywhere. I took bath and prayed for everything to go right. With light steps I moved towards my Lab. The clock showed 7:30. Fish was there in its aquarium, swimming happily. I was taking care of it for last four days and had developed liking for it. I didn’t want it to be sacrificed but that’s not what was expected from me. Aquarium was now covered with a sheet of polythene and nitrogen was introduced into it to reduce oxygen concentration of water. The oxygen concentration reduced to a level in two hours when my fish started to show signs of distress. This was the time. Lab had been meticulously cleaned to prevent any contamination. The fish was anesthetized and dissected. Then started two hour long protocol to extract RNA. I wasn’t allowed to use commercial kits for extraction which are easier to use. A master’s degree student in my Univ is expected to learn basics of each method and thus has to use old and elaborate methods. I was stressed out like a child appearing for an entrance exam. My throat would dry frequently.
Every step was being neatly done and just when I was about to do the last step I realized I had committed a blunder. I forgot to discard previous solution before adding the next one. This had dissolved all my nuclear pellets again. This just blew me off. I didn’t know what was happening then. I could see my fish cursing me for wasting its life. I didn’t want it. Still I continued keeping some composure. A step after another and finally a gel run to test RNA content.
With imaging system switched on the machines came to life. Lab had begun to buzz with people now and knowing that I had started my first experiment they were all excited too. So, the imaging system was switched on. Hearing beeps and cranking of the instrument my heart beat increased. It was the time that was to tell me if I was just a murderer of my fish. Gel was inserted and again the screen became blank before giving the final image. My eyes focused hard on the monitor to spot my gel. There were people around me to see my first research experiment giving its results. A flash again and the gel became visible. The bands shining clearly. A sigh of relief, few pats over my back and was into research, finally.