Another day.. another.. nickel?

So, I am taking a moment to breathe. My day thus far:

9am : Wake up (Yeah I am a lazy ass)
9:30 : After shower, check EQ2 auctions. Chat a few.. head in to the lab

10 am: Pull into Sonic and get my 44 oz iced tea, 2 Corn dogs, and some cheddar peppers (this is my brunch - yeah, I am REAL healthy)

10:15 am: Arrive at lab, bring food and books into office, run into my entire lab in the hall on the way out. They all ignore me, for the most part. Which is fine with me.

10:30 Eat, then head across campus to my lab building. (Office is in different building from my lab)

10:30-11am Start to prepare the reverse transcriptase PCR reaction for 20 samples from my chronically ill shrimp.



Get a phone call in the middle of this asking if I want to run another sequencing plate. I figure what the hell.. I need to dilute the samples and try to get better results anyway for another replicate.. so I say sure, I'll prep a plate "Real Quick".

Granted, it takes an hour to set up a plate:

Basically you make a master mix of some Really Expensive Stuff and add 1 microliter of each sample to a well in a plastic reaction plate.




Then you cook it in the cycler. It takes about 3.5 hrs. So mine will be done at about 6pm. *sigh*

Then I have to do the clean-up steps and load it onto the sequencer. At this rate I should be ready to do this part about 7pm. Might just load it tomorrow at this rate. Or they might entrust me with actually putting the plate in the machine All By Myself and pressing "Start".

So while this is cooking, I will continue with my Chronic RTPCR. Yay.

I'm STILL not writing. Guh. I guess I will write some after I start this on its way to cooking.

I've got 3 balls in the air today. At least it will go fast.

That seems so very time consuming. I admire your patience and ability to do the work. Hopefully it all works out in the end so your efforts are worthwhile! .... if it doesn't, then at least you know you can put up with mindnumbing tasks over and over for long periods of time. Sounds like you should run for presidency!

No kidding.. yeah it takes a certain temperament to do this stuff. We had one high school intern just leave in disgust after the first experiment that took about 4 days of tedious work fail and was told to "do it again".

I'm getting ready to inoculate 75 tubes from bacterial colonies. One thing I learned.. its far easier to see which are blue and which are white after they sit in the fridge for about 2 weeks.

If I get good yields, I think I will incorporate an "Incubate at 4 C for 2 weeks" step in my cloning protocol...

Hehe, two weeks? .... at least it gives you time to completely forget about it and be surprised if the results end up being a pleasant surprise.

Still not using Sigma's products huh! We make a well plate for them but it only has 96 holes!

Nice marketing Gavins!!!

*Hint* *Hint* Dori

Hehe, Dorina has the option to buy Sigma products already. I can't say theirs are better then the others though. We just run the tools to make the parts and they sell them.

Hehe.. do Sigma's plates work in Applied Biosystems DNA Analyzers? I use an ABI 3130. I'm using ABI's MicroAmp plates now, they tout them as specifically made for that damn machine.

I use Bio-Rad plates in the Bio-Rad Icycler for Real Time PCR. If their plates will work I'm all for it

Actually that plate does look like it might work. Its got the notch in the corner.

Dorina wrote:

Actually that plate does look like it might work. Its got the notch in the corner.

Im not sure what they work in, I know sigma designed their plate to fit in a certain machine. It also only has 96 wells, maybe I miss counted the one you have but thought it had more. *shrug* Its ok better you use something else then Sigmas and finding something wrong with it and bitching at me about it.