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Reverse transcription

From the LabLit short story series

Nik Papageorgiou 27 June 2011

All I need is a neat band to put in the paper. Instead, I’m like some fresh undergrad doing everything backwards

Friday 12:36 pm

Not bands in the gel; blue, nebular, tragic, leaking tears.

“Come on! Not again!”

Alex approaches the bench and peers over my shoulder. “Hmm. That doesn’t look good.”

I don’t have patience for his Zen attitude today. “Really. Thank you for your contribution.”

“Don’t get snappy. It’s just a Western blot.”

Just a Western blot? I need this yesterday.”

“For the paper?”

“Yes for the paper! The collaborators aren’t going to wait for me to sort out a simple protein detection.”

“What happens if you don’t make the deadline?”

“You know that equal contribution bit in the authors’ list?”


“I can kiss that good-bye.”

He shakes his head. “I see. Well, the day’s still young. Give it another shot.”

“It’s not like I have a choice. It’s just…”


“It’s just that I feel like, no matter how hard I work, everything keeps going backwards.”

Friday 12:21 pm

I sit at the bench and pipet on autopilot. The stink of TEMED fills the lab and the gentle cling-cling-cling of the magnetic stirrer mixing the gels drones away like a tribal instrument. Fifty microliters of this. Twenty-five microliters of that. Same method, same movements, same ritual.

And that’s not even what bugs me. It’s that when we sat down to discuss this Western blot, I said, “That? I can do that with my eyes closed.” Famous last words.

And now I look like the idiot postdoc who can’t pull a simple Western off.

Just one protein. 130 kilodaltons. All I need is a neat band to put in the paper. Instead, I’m like some fresh undergrad doing everything backwards.

Thursday 12:32 pm

My PI rubs the back of his neck and that little vein on his forehead quivers. “What do you mean it’s not working?”

“I think it’s the gel. It’s not setting homogeneously or something like that.”

He looks at the protocol I drew up. “Let’s see… Are you adding TEMED?”

“Of course I’m adding TEMED. It stinks up the place enough to know I’m adding it.”

“Okay, okay. So the proteins aren’t separating well?”

“The proteins aren’t separating, period. They just leak down all over the gel.”

He scratches his nose. “You know, the deadline –”

“Look at me. You think I need to be reminded of the deadline?”

“All right. Calm down. Let’s just go through the protocol.”

“I’ve gone through the protocol a hundred times.”

“Okay. Let’s go through it backwards.”

Wednesday 10:34 am

I stare at the clock on my desk. It’s a big break, this paper. Came out of nowhere like a Deus ex machina. Just share some of the FRET data and write up my method/results bit, and I’m second author. Catch the protein on a Western, and I’m equal contribution. Easy money, to quote a famous film.

Problem is, I’ve never done a Western blot in my life. There. Sue me. Is it my fault that both PhD and postdoc were consumed on bioassays, ELISAs and RT-PCRs? It’s not like you can shop for techniques out there. Still, it’s not something you tell generous, out-of-the-blue collaborators who want to put you on an Impact Factor 20 paper, is it? It’s like being a guy and admitting you don’t know the off-side rule. Awkward.

You learn. You draw up a protocol and you go through it, step-by-step. Don’t let the deadline pressurise you. First time it’ll probably flunk – but by the end of the second time, you’ll be able to do it backwards.

Tuesday 3:57 pm

Alex approaches the bench peers over my shoulder. “So, what does it look like?”

I don’t say anything; I just stare at the gel tank and try to ignore the flashes of light behind my eyes. I can feel the skin under my nose twitching.

He shakes his head. “That doesn’t look good.”

I blow out some air. “Alex, unless you have something helpful to –”

“How long are you setting the gel for?”


“How long are you leaving it to polymerise?”

I blink a little. “Uh, ten minutes.”

Alex raises an eyebrow. “Ten minutes? Where did you get that from?”

I shake my head and look at my miserable gel. “I don’t remember. Some paper or something. Why?”

“Because I’d say that in this lab’s ‘room temperature’, thirty minutes should be your minimum.”

My voice shakes a little. “Really?”

He passes me an Acrylamide bottle. “Yeah. Really.”

“Thirty minutes?”

“Here, use my timer. Pour your gel and count thirty minutes backwards.”

Monday 12:36 pm

“Oh, come on! Not again!”

Alex looks up from his bench. “What is it?”

“The proteins are leaking all over the gel.”

He comes over and he peers over my shoulder and for some reasons I have the strangest feeling of déjà vu.

Alex pulls his Zen face. “Hmm. That –”

“– doesn’t look good. Yeah, I know.”

He smiles. “We finish each other’s sentences. Been here too long. Is this the Western blot for that paper you were telling me about?”


“Well, don’t get discouraged. Try it again.”

Again? I need this yesterday!”

“Don’t get snappy. The day’s still young. Give it another shot.”

He turns to leave and I realise my timer has stopped working. Battery died.

“Hey Alex?”


“Sorry I snapped. It’s the stress.”

“It’s okay. Time’s against you.”

“Yeah – well, not just that. It’s just…”


“It’s just that I feel like, no matter how hard I work, everything keeps going backwards.”