Wednesday, June 09, 2004
Should have stayed in bed
I think everything that could possibly go wrong today, did.
Now that I’m an “expert” at the 2D gel protocol, I decided to go all the way and add the second digest, which will tell me which direction the replication fork is coming from. This means that after I cut out the first dimension, I have to soak the slices in different buffers several times for up to an hour for each wash. Then I ad the enzyme (onto one of the two slices, so I have a control) and let it sit for 4 hours at 37 degrees. And THAT meant that I had to drive my ass BACK to the lab at 11:30 last night and work until 1 setting up the second dimension.
But it’ll be worth it, right?
This morning I come in to check my gel. Something doesn’t look right. Perhaps that’s because all the buffer that’s SUPPOSED to be in the gel box is now everywhere but. The tubing in the pump burst. Shit. My gel looked OK (the leak must have happened about an hour or so after I had started the gel) , but I had to run around trying to find new tubing to cram into the pump. Found new tubing, got everything set up again, thinking I can run it at max voltage all day, come in again tonight, and set up the blot.
After about two hours, I checked the setup. The gel was running fine, but the pump was not. The pump refused to work at all. The pump had blown a fuse.
To make this quick, I got the fuse replaced, but decided to use another lab’s pump until we can get the proper tubing for ours (I’d just been using regular tubing, but you’re supposed to use expensive MASTERFLEX tubing. at $78 for 50ft, it’d DAMN WELL BETTER WORK!!!)
It wouldn’t have been nearly so stressful today, except that I started another round of DNA isolation, and needed to get the CsCl gradient set up and in the ultracentrifuge tonight. In the confusion of it all, I think I may have used NIB buffer instead of TEN buffer. Shit. I hope it still works!!! I don’t think my nerves can take anymore today.
PS: I think I may be the only blog out there that mixes relationship/sex chat with hardcore science. And I’m not sure that either of those subjects is of ANY interest to anyone but me. Oh well. It’s the only therapy I can afford right now!
// ranted by evilsciencechick @ 6/09/2004 02:57:00 PM
Now that I’m an “expert” at the 2D gel protocol, I decided to go all the way and add the second digest, which will tell me which direction the replication fork is coming from. This means that after I cut out the first dimension, I have to soak the slices in different buffers several times for up to an hour for each wash. Then I ad the enzyme (onto one of the two slices, so I have a control) and let it sit for 4 hours at 37 degrees. And THAT meant that I had to drive my ass BACK to the lab at 11:30 last night and work until 1 setting up the second dimension.
But it’ll be worth it, right?
This morning I come in to check my gel. Something doesn’t look right. Perhaps that’s because all the buffer that’s SUPPOSED to be in the gel box is now everywhere but. The tubing in the pump burst. Shit. My gel looked OK (the leak must have happened about an hour or so after I had started the gel) , but I had to run around trying to find new tubing to cram into the pump. Found new tubing, got everything set up again, thinking I can run it at max voltage all day, come in again tonight, and set up the blot.
After about two hours, I checked the setup. The gel was running fine, but the pump was not. The pump refused to work at all. The pump had blown a fuse.
To make this quick, I got the fuse replaced, but decided to use another lab’s pump until we can get the proper tubing for ours (I’d just been using regular tubing, but you’re supposed to use expensive MASTERFLEX tubing. at $78 for 50ft, it’d DAMN WELL BETTER WORK!!!)
It wouldn’t have been nearly so stressful today, except that I started another round of DNA isolation, and needed to get the CsCl gradient set up and in the ultracentrifuge tonight. In the confusion of it all, I think I may have used NIB buffer instead of TEN buffer. Shit. I hope it still works!!! I don’t think my nerves can take anymore today.
PS: I think I may be the only blog out there that mixes relationship/sex chat with hardcore science. And I’m not sure that either of those subjects is of ANY interest to anyone but me. Oh well. It’s the only therapy I can afford right now!
