The WORKSHOP ON ANALYTICAL TECHNIQUES IN GENOMICS AND PROTEOMICS started yesterday with 61 interested undergraduate students! This workshop is conducted by the Department in association with Central Dogma Pvt. Ltd. Pune. Details of the workshop can be found here!
The workshop is a 4 days hands on training session that includes various interesting experiments as follows:
- Plasmid DNA Extraction
- Genomic DNA Extraction
- Agarose Gel Electrophoresis
- PCR and Primer Design
- Plasmid mapping using Multiple Restriction Enzymes
- SDS page gel and so on
Yesterday, 11th April 2016 was the first day of the workshop and we covered the first three experiments from the list above. It was one continuous experiment that covered all the three points.
Being a Second year undergraduate student, I knew the experiment in theory to a certain extent, but had not performed it practically yet!
Gel electrophoresis is a method for separation and analysis of macromolecules (DNA, RNA and proteins) and their fragments, based on their size and charge.
Before I talk of my wonderful experience during the session, The snapshot summary of the experiment is as follows:
We were given 6ml of culture from which we would extract the plasmid. We centrifuged and re-suspended the pellet in three buffers, one at a time; namely – P1, P2 & N3, each having its own function. With the help of a spin column, we then centrifuged the pellet by giving it a PE (Ethanol wash) to remove any unwanted traces, We then gave it a dry spin and finally eluted it in a fresh eppendorff tube. This time we did not discard the supernatant but the contents of the spin column. The supernatant contained the plasmid. Now that we had our plasmid, we loaded it onto our Agarose gel plate.
The instrument on which the Gel Electrophoresis experiment is performed contains of three parts, namely – the Buffer tank, the Casting Tray (Gel Plate) & the Comb.
The Buffer tank is a deep rectangle, box like structure wherein the Buffer is added.
The Casting tray is a flat tray like structure where the gel is added, its free ends are sealed with a tape to ensure that the gel does not leak.
The Comb, also know as Well Comb is a bristle bearing structure and is used to bore wells in the gel.
Here’s an image of the same:
Once the wells are formed, the supernatant that contains the extracted plasmid is poured into the wells.
This is then connected to the voltage supply where the gel is made to run, obtaining two dyes- blue & violet.
And then this gel is observed under UV light, where the extracted plasmid could be observed. Given below is an image of the gel under UV transilluminator.
The light fluorescent orange bands between the violet and blue dye is the Plasmid.
The experience of this session was fascinating!! We learnt many new things and had a great experience!!
The hands-on experience was so good especially gaining it at an undergraduate level. We used micro pipettes for the first time, we learnt how a split column with its collection tube functions.
We also used those super cute eppendorff tubes!!
Anjali Ma’am, founder of Central Dogma Pvt Ltd and her team were very patient and were more than happy to explain us the concepts till we got them correct.
This workshop is excellent as it is helping us to still keep up with our academics even during the vacation! And we are having such fun through the amazing practical work that is designed, that I think hardly anyone would refer to this as work!
Looking forward to today’s session!! 🙂
Draft by: Lisha Ramani