Upasana Simha

Upasana Simha 3rd Year student, Srishti school of art design and technology.

An interest in bio and a need to explore an unknown space is the reason I am here. The work we're going to now be doing is so different from what I am normally accustomed to working with, and the fact that I am not from a science background throws me into a space with no limits or bounds to work with.

DNA

15 micro litre of water

E.coli cells

Ice

LB(lurea Burtani)

Centrifuge machine

Process: -take 100 micro litres of the E.cole cells along with 3 micro litres of the plasmid.

-Incubate in ice for 40 mins.

-Heat shock is given for 90 secs at 42C

-It is then put in ice for 2-5 mins.

-You grow in 990 micro litre of LB for 1 hr at 37C.

-centrifuge for 3 mins at 8000 rpm

-we then discard 900 micro litres of the supernatant

-There is a pellet remaining which we then dissolve in the remaining 100 micro litre solution.

-We then prepare Lb agar plates which contain 100 microfram/ml of ampicillin.

-Incubate the plates at 37C overnight.

HANDS ON!!

Its great that we are actually getting down to doing some work in the lab. My understanding and approach to these processes are changing and its finally getting clearer. The association of this proces aoriented work was related to cooking by someone, and to a certain extent the comparison is valid, but the patience that one requires to be able to work in a lab is a skill in itself.

One of the most important things I learnt is how it is essential to always be aware of what you are doing at all points of time. One small mistake at any point in the course of your experiment can cause havoc.(An experiment that should ideally take 2-3 hrs long can sometimes take 2-3 months to complete!!)

Working in this environment has made me realise that this is all happening, that we are currently no longer in a space we have no clue about, because we are finally getting an understanding by actually getting in there..

WHAT DO WE KNOW WE'RE JUST DESIGNERS...!!

Transformation, plasmid preps, digestion and ligation ask me the process and I'll be able to tell you whwn to add ALS 1 and how long you need to centrifuge and at what temperature you need to incubate the bacteria. Hearing all of this I felt like I knew all that I need to know to grow my own bacteria. Indeed I was truly proud on our progress and how swiftly we had adapted to this environment that spoke a whole new and different language from what we understood. As I was saying I was so amazed at this technical language That i felt it was a must that i show off my newly aquired skill, one such time that I was yacking away the process this one person just looked at me very simply said "so why again do you do the plasmid prep?" I scoffed at the question and thought it was the silliest question ever. I mean a person who can rattle of the process obviously knows the need for a plasmid prep right? WRONG!!!! i had no clue, this question had me stumped. I realised that I may be able to tell you how to grow your own bacteria, but I can give you no logic behind the process. I felt like the designer(the outsider) all over again. This is when another realization dawned upon me, I am always going to be the designeer, my learning the way I was going about it was superficial. Maybe it was always going to be a superficial learning. The way we can possibly try and get a better understanding is by asking a lot more questions and learning why and how something is happening. Now not only do I know that I shouldn't go around babbling about my bacteria experiments, but also know a little something about how and why we're doing a particular step during the course of an experiment.

MY VERY OWN SCIENCE THEORIES 

PLS NOTE THE THEORY IS COMPLETELY AND ENTIRELY WRONG.

This was when we didn't fully understand why and how certain things were being done in the lab. Before we could actually get to the lab and find out, a bunch of us sat together and tried figuring out why we were performing the plasmid prep. This is what we came up with.

In the bacterial transformation we grew colonies of bacteria which possessed thelysis gene and the pbad gene. While we were looking up theses processes we happened to come across this line that said a portion of the DNA cant replicate, it needs to be inserted within a plasmid in order for it to multiply. Therefore we came to the conclusion that th bacteria we grew could not replicate to produce future generations of bacteria containing the lysis gene or the pbad gene. Which is why we move onto the next step-plasmid prep. This step is meant to grow plasmids into which we can insert our gene, such that it can replicate to produce bacteria babies containing the genes we want.