Failed experiments but undefeated

What seems to be straightforward in theory turned out to be full of obstacles in practice.

Theoretically, ligation sounds simple where you simply add a vector (a carrier), an insert (your gene of interest) and a DNA ligase (an enzyme molecule which join two pieces of DNA together) in a buffer mixture, and incubate at certain temperature for a period of time.

With this mentality, I performed two ligation experiments but they did not gave result. I was frustrated because I preconceived that it should work for a simple experiment and I am under time constraint. I was determined to get it right!

Coincidentally, a visiting professor told me that some labs were having problems ligating and after discussing with my friend, Saimun over Skype, he corroborated the difficulties involved to optimize many parameters for a successful ligation. He also suggested his ligation recipe. These helped to alleviate my frustration where I thought I was bad at ligation.

With each experiment, I learnt more about ligation and thank God for giving me a sharp observation, I troubleshot the possible mistakes which I committed. The two main causes could be low concentrations of my inserts and short incubation time.

At the third attempt, there were results which needed to be verified next week. For now, I am partially satisfied till I get the verification. I do not like to count my chicken before they are hatched.

-----------------------------------------------------------------------------
Lessons learnt from this mini-episode.

Mistakes are meant to be made because we are not perfect and we can learn more through mistakes than success.

Failure is a state of the mind; When you succumb to it, you are defeated.

Shortcuts end up being nowhere.

Be humble to seek help.

Learn from the experienced and adapt their advice to your situation since you are walking it, not them.