Waste not, want not
My intrepid benchmate had a bit of a rough time today. Nothing, she lamented, seemed to be working. She came in to an incubator full of agar plates with nothing growing on them, which apparently set the tone for the rest of the day, unfolding as it did with various minor disasters and pesky obstacles. Nothing devastating — just enough to annoy.
But she was most concerned about her blank plates: seven entirely different supercoiled plasmids, all zapped into expensive competent bugs from a reputable company starting with S. What, she speculated, might have gone wrong? We talked through her protocol: the bugs were working fine, because I’d used them myself only a few days ago. Her transformation manipulations were text-book, and the DNA itself was above reproach. We were both stumped.
But then she slowly reached for her lab notebook and muttered, “Hang on a minute.” A few moments of page shuffling and plate examination ensued. And then:
“Oh, damn.”
Somehow, she’d plated the two kanamycin-resistant bugs on ampicillin plates, and the five ampicillin-resistant bugs on kanamycin.
Ouch.
But fortunately, there was a happy ending. She realized that the tubes full of bugs were still somewhere in the bin underneath our bench. Scrabbling through the trash, gloves and pipettes flying every which way, she finally managed to find all seven tubes and was able to restreak all of her transformed cultures onto fresh plates. (I told her if she really wanted to go for it, she should re-use the plates that nothing had grown on. But she wasn’t too impressed with that idea.)
The purists may frown, but I’ll bet they grow. Stay tuned.
Ha ha! I’ve done that! And recovered it the same way.
I don’t recall being quite brave enough to re-use the plates though…
I’m a purist. I’m frowning. Ha! can’t even write that with a straight face.
Bugs should grow fine. Not like wimpy mammalian cells – “oooh, no, I’ve contacted air outside the lamina hood oooh noooo I’m contaminated ooooh noooo.”
Bugs know no fear. Bugs are survivors.
Lots of juicy colonies this morning!
Hurrah.
Huzzah!
Been there, done that, but unfortunately never came across the recovery method in any paper.
There should be a rule in all labs: wait 24 hours before getting rid of your rubbish, you just might need it…
Ha! That was my first major mistake in lab… I thought the E coli strain i was using was amp resistant… It wasn’t– Note: ampicillin kills E coli– my labmates havent let me forget that one!
In a previous postdoc as a microbiologist working on virulence determinant regulation in S. aureus I did this kind of strain/plate mix up thing to the extent that I knew the end of your story almost straight away! I have also used the wrongly used plate again, but that depended on the kind of experiment that I was doing. It does work.
Furthermore, I once had a troublesome ligation that I was doing, and eventually solved it by finding a tube of a ligation experiment that I had lost the week before. I found it on the floor whilst I was looking for something else I had dropped, and I thought I may as well give it a go as I still hadn’t managed to achieve my cloning. As I say, it worked, but I didn’t write it up in my lab book as “lost tube, kicked it around the floor in the dust for a week before rediscovering and transforming into competent cells” That just seemed wrong!
Wow–week-long ligation? That’s *worth* writing up!