I have no idea whether to say I’m lucky or unfortunate… haha!
After dozing off while waiting for the sonicator with it producing tonnes of whistling sound, pulsing all the way, I decided to take a break while I have free time. So, after placing the bottles in the centrifuge, I decided to sneak off for a break with some nice lunch, at least. Summer shared two songs by Eason with me, which I have no idea what’s the name but heard before. And then, I hopped on the bus and went for Japanese food at Arts Canteen. I met my friend over there and we chatted. After she left, I propped open my log book to record down what I’ve done yesterday and while I was writing down the buffer I made, something came right BANG!
Yes, it’s a WHAM! I realized the urea I added for my denaturing condition buffers are not the correct ones!!! Looking at the manual of ‘8M’, I checked and found ‘8mM’ in my calculation on C-fold towel. The first thing I reacted was a ‘dang’ in my head. How could I be more careless than this?! And then I remembered that while I was preparing the buffer, I was supposed to adjust the pH using HCl and when I checked mine, it is below 7.0, which means I used NaOH. And I remembered I used up the whole bottle of NaOH. If stupidity and careless are crimes, I will be the number One Wanted person now! I should have notice something is wrong when I have to use NaOH instead of HCl!!! And I was wondering did I added the correct ingredient! It was not the type of ingredient, it is urea, the one that gives alkaline numbers, that is insufficient!!! And Aldo reminded me before that the problem with urea-based buffers is that it is very difficult to dissolve. And yesterday I was so happily preparing the ingredients without any difficulties! I should have sensed the weirdness!!!
Anyway, yes, I recalculated the figures on the spot and travel back to lab just to redo the buffers. Ah, but then I realized how difficult it is to dissolve those seemingly lovely urea balls. But nonetheless, I shall consider myself lucky instead. Instead of realizing how wrong I am when I need the buffer or AFTER I used it, I found out about it before all these went wrong and affect my experiment, lucky, really! Imagine I put all the buffers in and realize it was not working just to find out that the urea content is not right in my buffer! That would be much more cruel, I daresay…
Yeap, this is an incident to remind myself to be much more careful next time… no more mistakenly omitting a milli, haha! Wish everything will be okay and yes, I still have half a bottle of urea balls to dissolve tomorrow with two buffers to be adjusted back to the correct pH. These are interesting events… not to mention being lectured by the lab officer because our lab is suddenly short of those blue-capped bottles… Okay, shall stop here – Research is not easy, be it in Science or in Arts .