You know, I think I may have fried my brain cells or something. Lately, I've been doing many stupid things.
It must have been my incompetence, as I seem to be multitasking most of the time. Take last Friday for example, I went to lab very early to do vector control and to run (the umpteenth) genomic DNA extraction at the same time. Kind of stupid, really, to run two experiments together, but I was greedy and didn't want to waste a second of the waiting time. To add stress to it, I had to finish transforming ligated DNA and extracting gDNA both by lunchtime as I had a career workshop after lunch. Now, anyone experienced will laugh and tell you it is impossible to complete so much work in half the day. But I was greedy, laid out the materials on two opposing benches and spent the entire time going back and forth from one bench to another. On one bench the centrifuge is done spinning and keeps beeping, on the another bench the digested mixtures are completed awaiting for gel electrophoresis. Gyaa.... what a dizzying exercise.
In the end, I couldn't finish the experiments in time and was late for my workshop. My mentor had kindly run my DNA extraction gel electrophoresis for me, as I skipped lunch to rush to the workshop. After the workshop, my mentor told me my DNAs were weird. The extraction yield was very good, but there were EXTRA bands. I was like, "nooo.... my whole afternoon is wasted again!!" I quickly went back to lab to troubleshoot and re-run the gel using one of the 16 samples, and found no extra bands O.O The DNA must be playing hide-and-seek with me.
However, I still could not leave as my transformation part of the experiment was not completed and I had to wait like an hour for the bacteria to grow before I can spread them on antibiotics plates for overnight incubation. When I went to the cold room to get plates, I realized there were NO MORE PLATES!!!! The antibiotic plates I wanted were not in stock!! Talk about bad luck! T_T What were left were a stack of plates labeled "Not-so-well-made-Amp-plates".
-_-''' I do not know what the person who made the plates meant.
I had no choice but to use them anyway, and because these plates were 'defective', I had to plate on another type of plate which were good quality, but had slightly different nutrient composition. Meaning, I had to spread TWO sets of plates. Now, you might think, another extra set only mah, what's the big deal. It means I had to spread 13 samples on 52 plates. And that time was already 930pm.
By the time the bacteria had happily grown to the state I wanted them to, I was the only person left in the level again. T_T I don't really enjoy OT, because the quiet lab can be quite creepy! I think it must have really just been me, but I keep hearing sounds. However, thanks to that, I have learnt some plating-acrobatics, as I could now spread plates without even looking at the plates (half the time my eyes were averted nervously to see if any vampire, uh I meant person, sneaked up behind me).
Then yesterday, I went back to lab to take out the incubated plates and do genomic DNA extraction A.G.A.I.N. (when can I get enough DNA??) Fortunately, my vector control went well; meaning I can proceed to actually start making the molecular library. After finishing my DNA extraction, I run all my 32 samples on gel to check the DNA concentration. I had purposely made brand new gel from clean TBE liquid and ultra-grade agarose, so I was confident my gel quality was good. Furthermore, I had performed the extraction using the conditions I optimized last week, so I was quite sure my yield would be pretty good.
So, I loaded my samples, covered the tanks, and plug in the cables. As I was connecting the cables, I noticed the tank orientation was upside-down. "But it's okay, as long as I plug the electric poles right," I thought. After running the gel for 45mins, I took them out of the tank for EB staining. As I did, I thought something was weird. Usually, the blue dye from the samples can be seen at the end of the gel after electrophoresis, but I didn't see them on my gel. "Maybe my gel's a bit too short, but it's okay, since the bands are not very low," I had thought. So, without further thoughts, I put both pieces of gel in for staining... and waited 25mins for viewing.
After staining was done, I took the gels to view under UV. What I saw on the computer nearly made my heart stopped beating.
NO BANDS.
When I viewed one gel which had two rows of wells, I noticed something odd: the bands from the lower row appeared on the upper row gel. At first I thought I placed the gel upside-down on the platform. But it was quite impossible. because the ladder was on the correct side!
Then, it dawn upon me.
I had ran the gel BACKWARDS.
The electrical poles were right, but the gel was oriented upside-down. Thus, all the DNA from the upper row were run into the tank liquid. How stupid can it get?? I didn't know whether to laugh or cry when I saw the gel photo. I mean, WHO RUNS GEL BACKWARDS??
I gave up for that day. Maybe it's because my concentration level had hit low and I'm beginning to do very unsensible things.
Lesson is.... always keep the tank on the right orientation.
～みはる～
By
15 years ago
