Had a really bad day today... I would say that today was shitty for the lack of better words. Why? I'll make it simple and to the point. Only one person can get me so frustrated at the labs... Yeah, the one and only K. I will withhold all names for obvious reasons. I'm pretty lucky that I'm not handling this subject alone, in a way. KS was a lot of help today. Now, where should I begin? Right, the beginning it shall be.
The problems actually started yesterday. K asked me to prepare 4 bottles broth of the morning session and 3 bottles for the afternoon session. Note, each flask holds 50ml of broth, which means a total of about 350ml. The practical states that each group is supposed to use 10ml of bacterial culture, which means a total of around 140ml for the morning session alone. The day I received these instructions was Friday. I made the solutions on Monday and I began to prepare for today's practical on Tuesday. On Tuesday, I had to aliquot the 30% glycerol into 28 tubes for the morning session and 26 for the afternoon session. I managed to set up all the things for the practical by 5.00pm and was getting ready to leave for the day. It was then that I realised that K liked to have at least 2 samples for each group. I hurried to the practical steps and re-calculated the total volume of broth I needed. In total, I needed some 600ml of broth. The worst thing is that the broth would require autoclaving to make sure there was no contamination. I needed to be sure so I called K up to make sure. Sadly, my worries were no unfounded.
KS and I quickly whipped up 3 more bottles of broth and called it a day. I would need to come in early the next morning to get the competent cells ready so I decided to autoclave the remaining 3 bottles the next morning after I added the inoculants into the available bottles. So, this morning, I rushed to the labs and by 8am, the next batch of cells was multiplying in the shaking incubator. By 8.30am, I managed to autoclave the remaining 3 bottles, grab heaps of ice, set up enough calcium chloride to drown a rat, and distribute chilled 30% glycerol to the classes. By 9 am, I was checking the OD of the cells in the incubator. The increase was slow and I was worried that the OD would not rise up quick enough for the class. Around that time, KS came in to work.
By 9.10am, the lab was bustling with students. K's wife is the demonstrator for one of the lab classes and personally, she's as annoying as K himself. She has this knack of being altogether condescending and asking for things when I'm busy. The worst thing is that I have to drop whatever I'm doing to cater to whatever inane thing that she's asking for. That, however, will be a tale for another day. By 10 am, K was getting antsy over the OD of the cells and asked me to add 1ml extra bacterial culture to the fresh batches. I responded quickly and began to fill in pipette tip boxes. It's really amazing how quickly students go through them!
Around 10.30am, K couldn't wait any longer and took the new batch out and began to distribute the cells to the students. I don't think he realised the extra 2 bottles that had to be used. I was just beginning to think that I could sit down and breathe when he popped in to tell me the amount of glycerol I provided was insufficient. At first, I thought he didn't notice the extra tubes I added in the ice box. Sadly, that wasn't the case, as the final volume of the cells would be around 1ml, the students would be able to make 10 replicates. 10 replicates would mean that each group would produce 20 tubes of competent cells. In the semesters before, only 2 replicates were used and I followed the instructions left by the previous technician before me. I had only prepared for the students to have only 2 replicates and suddenly, I had to make more glycerol.
At that moment, I felt like shit. I was so frustrated with K that I was very close to crying. After regaining my composure, I calculated the amount I needed in total and did the math to determine the volume of stock solution I needed to achieve the 30% concentration needed for the practical. I was feeling rather rubbish-y as the glycerol was supposed to sterile and I didn't have time to autoclave the thing. K suggested I just filter sterilise the solutions, but that wasn't the case. The case was the fact that he ALWAYS INFORMS ME ABOUT THINGS AT THE VERY LAST MINUTE!!!!! This was the main reason the previous two technicians left. They were tired of him making their lives so difficult when working under him. One of them used to stay very late, the other used to come to work even on weekends when the results could not be achieved.
I have noticed a trend though. Every Wednesday, I will definitely have problems in the morning sessions. By the time it's afternoon, I'd have covered everything and made sure that K wouldn't need to bother me. Oh well. That's my rant for today at least. I need to sleep... I've been rushing to and fro at work so much that I am drained. Toodles...
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