Remember my cloning dilemma story? Well, it slowly turned into a nightmare.
I made my way back to campus to continue working on my research. It takes time to readjust to everything I left behind for two weeks, hoping that this time, things would finally fall into place after all my previous cloning attempts.
Coincidentally, the research supplies I had been ordering — new ligation kit, primers for colony PCR, and a few other essentials, had finally arrived from the supplier.
Everything was ready.
The materials were there.
The experiment was waiting.
So, I decided to give my ligation mixture another shot, this time using the Quick Ligase from NEB. Everything seemed great. It was quick, just like it was named for — Quick Ligase. It saved me a lot of time, and for once, I thought maybe this cloning experiment was finally going to give me some hope...
The ligation was ready.
The samples were ready.
I was ready.
When I was about to move on to electroporation, I switched on the electroporator…
And then — ERR. ERROR. No beep. No warning.
At that point, I was hoping it was just a minor technical issue. Spoiler alert: it wasn’t.
Yup. The electroporator decided that my cloning experiment was the perfect time to have a problem. And just like that, my supposedly “quick” ligation experiment came to a very un-quick halt.
So, I did what any researcher would do when things suddenly go wrong. Apparently, one of the most important skills in research is learning how to search for answers at 2 am.
1) I went down the rabbit hole of Google and ResearchGate
I tried to find out whether other researchers had experienced the same problem — and to convince myself that someone, somewhere, had already suffered through exactly the same thing.
2) Repaired by my own
What was I thinking? I'm not an engineer or a technician.
3) I contacted the supplier
I reported that faulty electroporator error and asked for their advice.
4) I reported (cried) everything to my supervisor
I started thinking, maybe I should just start all over again using the conventional heat-shock method.
Back to basics. No electroporation. No fancy troubleshooting. Just my competent cells, my quick ligation mixture, and a little bit of hope.
But guess what?
The refrigerated centrifuge was at faulty too.
One instrument broke down, then another. Every time I thought I had found an alternative route, something else decided to stand in my way.
It may not be going according to plan. Instead, I somehow ended up sending away that faulty electroporator for a diagnostic repair and it definitely took longer than I expected.
But hey, better late than never.
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