First DNA extraction!!!

First DNA extraction!

DNA bands observed using UV light.

It's been a long time since I wrote something out here, was really caught up with studies and assignments, But now that we are facing a global pandemic COVID-19 and everyones' quarantined, I've got some time to spare.

I hope this COVID-19 gets over soon (*fingers crossed*).

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It was the month of September. We had an experiment in biochemistry, wherein we had to extract DNA from plant source using chemicals. Sound pretty simple eh? But can y'all imagine we remove the DNA like "legit" the DNA from the cells which we can't even see with our naked eyes!! Chemicals play a super duper important role out here!!!

We had a protocol which we followed. Following the protocol is an easy task to do in comparison to  preparing the reagents. Naming two-

Buffers- making buffers was a task! specially when it comes to maintaining pH!! (if it goes to acidic bring it back to basic and vice versa!). First we used pH papers to maintain it and  later on pH meter (accurate readings). Takes nearly 15-30 minutes! 

Why do we have to use buffers in the first place?... because it helps maintain the pH. As we have to break open the cells to extract DNA  (chemicals break cells open)  a lot of components remain suspended in the sample so to neutralize it we use buffers. 

For detailed protocol check- Sambrook & Russell (2001). Molecular Cloning: A Laboratory Manual (4th edition) Cold Spring Harbor Laboratory Press.

The DNA sample (from plant source) is treated with chemicals until one obtains a solid pellet of spooled DNA. The amount obtained is crazy small (micrograms)

The pellet at the bottom is DNA.

Casting the gel-  After you obtain your DNA sample in pellet it is suspended in a buffer again and stain is added to help visualization.  Casting a gel simply means running of DNA bands on Jelly like chemical (Agarose) under the influence of electrical field.

It takes 15-25 mins for the DNA band to move across. Our professor is pretty cool, till the gel was running he gave us a "Chai" break!

Right hand side the samples are loaded which will move to left side. (Negative charged DNA moves to the positive terminal)

And then there was joy. And then there was DNA atlast!!!!!

DNA bands observed under UV light.