Guess who is stuck in lab on a Saturday with the cheering prospect of being back on Sunday?
For the past few days, i’ve been super distressed because the cloning I was doing didn’t work. No reason. Just didn’t work. For the life of me, i couldn’t figure out what i did wrong. So i said “Screw it. Imma gonna redo the goddamn thing.” Hence the weekend dash to the lab. Thankfully, i still had prepped-DNA lying about, all digested, purified, and ready to be glued together. Saves me a lot of time.
Step 1 was to glue together the gene I want with some other DNA in a tightly coiled ring. I got the DNA-gluing step going last night. Then I went out and got drunk. I came back to lab in the morning to finish Step 2. This step involves coercing bacteria into accepting my gift of knitted together DNA. My gift makes them resistant to ampicillin. A gift that keeps on giving I tell you! and yet, I must heat shock them into accepting! Step 3 is to take these cells carrying the extra DNA i added and spread them out on plated of agar so i can get nice individual colonies of cells. The agar plates contain ampicillin. This way, only the cells that have my DNA in them will grow (because they are resistant) and everything else won’t. 😀 That, in short, is the process. If it works, i’ll be VERY happy. If it doesn’t, i’ll be less happy. I’ll have to repeat everything! Not just these three steps, but all the steps leading to these three steps… That’s almost a week’s worth of work.
Anyway, while my media plates were being prepped, i decided to read random blogposts to while away the time. And the very first post i landed on was this one. This blogger’s colleague had the same problem as me. Transformation didn’t work. And she DID find a solution to why not. Her solution won’t work for me though, because I’m positive i’m using ampicillin plates for ampicillin resistant bugs. I checked. Thrice. But it comforts me, sitting alone in lab on a saturday night, to know that there exist other researchers in the world who have technical problems similar to mine. Much like this xkcd comic.
Here’s hoping my experiment works this time. If it doesn’t, i’ll cry.
OK, maybe a little.
Also, you really should consider heading over to my #Scifund project page and helping out. Much obliged.