The Curious Case of the Ghost Blot- Halloween edition

The dark room was extra eerie that Friday evening. It was late and most of the lab members had left for the weekend. Neither the sharp tang of developing fluid nor the noisy whirring of the Imaging machine offered much in terms of company. But company was the last thing on the mind of the sole occupant of the dark room. A little 3”x4” image was due to appear on the computer screen any minute now. The 10 second wait prompted a train of thoughts, in specific order:

  1. Did my experiment work? What if it didn’t work?
  2. I guess I could repeat it… that means working over the weekend. That means no Doctor Who binge watching… not ideal, but not the worst situation.
  3. What if the repeat doesn’t work? Perhaps I made a major mistake somewhere along the line…Its coming back to me now! I used 10ml of reagent A instead of 8ml this morning! How bright am I!!?
  4. I deserve to miss my Doctor Who marathon.
  5. I’m so hungry right now.
  6. No wait, I did use 10ml of reagent A. Not so dumb after all am I (*imaginary pat on the back*)
  7. What if all of my experiments fail (loop to thought 1)?

As the whirring comes to an abrupt halt, one could almost hear the machine say ‘Boo!’.  A little image appears in the screen. It is a blank image- the ultimate Western Blot nightmare. Now that can’t be right (unless, refer to thought 3).

But wait, the contrast button has been turned off. Must have been the *expletive* who used the machine before me. A quick click of the Contrast button and right there on the screen were three perfect ‘bands’. Did I actually do something right? Did my experiment actually work? The last 6 months of work were not in vain after all! The temptation to frame the image and hang it in my living room is high, but I resist. Not because I’m a sensible adult but cos of copyright issues. Bummer.

So what is so scary about a little 3”x4” image?? Lets ponder….

A typical scientific research project works like this :

– Identify a question. Eg- Does gene X cause cancer? Will treating patients with drug Y reduce  their cancer?

– Figure out how best to answer the question. Eg- What experiments would best answer my question. How much time and resources will this experiment cost? Is it worth it?

– Design the actual experiment- order stuff needed to do the experiment

And finally – Do the actual experiment!

Behold, the ghost blot…
(although generally the bands are less bat like)

There are some methods (often called ‘assays’) that help us find out if the experiment actually worked. The little 3”x4” image is an example of one such assay called Western Blot*. In the above (admittedly dramatic) narrative, I used the Western Blot assay to check if my experiment worked. Because the whole process (as straightforward as it looks) takes up time, energy, brainpower and resources, the prospect of it failing can be quite scary.

Western Blot is one of many assays that are used by molecular biologists to make sure that their discoveries are reliable and that some (if not all) can perhaps be used to benefit patients in the future. Which is why assays are repeated and perfected till they are absolutely reliable. The process itself might appear to be complicated and tricky. But at the end of the day, research is about proving your point. And the joy of being correct makes it all worth it!

Happy Halloween to all my fellow Western Blotters… And many more ghost blots to come!

* What is Western Blot?-

Written by Sathya Muralidhar, creative input for this blog was contributed by Dr. Juliette Randerson-Moor