Who ever invented rope is a total a-hole

What do you do when you been spending every day for the past two weeks in the lab (for 4-9 hours a day)? Well if you are me, you go see the movie "Blades of Glory" with Will Ferrell and Jon Heder. I knew that the movie was going to be really silly but honestly it was totally what I needed. It was nice to just relax and not really think about anything for two hours. It was pretty enjoyable and a good break (also the source of the title). And I am feeling pretty good because I figured out the NPR Sunday Morning puzzle almost right after I heard it (unlike usual where I have no idea what the answer is). There are also two episodes of "King of the Hill" tonight making today one pretty good Sunday.

"Wait a minute. Every time I leave, you call John Redcorn. I know what's going on here. Your headaches are a desperate bid for my attention. But what do I do? I pawn you off on some Indian healer so I can have my Dale time at the gun club, or breeding show turtles, or on the Internet investigating unexplained phenomenona. God, I am so selfish!" - Dale Gribble

Slowly collapse like a flan in the cupboard

Things in the lab have been going pretty well, but very busy. I went in for about 8 hours over the weekend, which actually wasn't that bad because there wasn't as many people in the lab so fewer people asking me about 30 questions each and trying to share equipment. Which was one of the problems that I had today. There is a labmate that I mentioned previously who is difficult to work with mostly because she feels that her work takes precedence over everyone else's work. Everyone else that I share lab space with tries to work together so everyone can use the equipment and space with as few problems as possible. The equipment that everyone was sharing (about 5 people) today was the centrifuge. At about 3 pm, the bitchy girl comes in and claims that everyone using the centrifuge was getting in the way because her experiment is time sensitive. It would be logically for her to tell us in the lab that she would need the centrifuge at regular intervals so we could plan around it (since we were already planning around 5 people). By the way, the girl never comes in on the weekend and leaves without fail at 5pm. So it is any wonder that working on the weekend is more enjoyable?

DALE: I have more pressures than any of you. You ever try replacing a cockroach's blood with root beer?
HANK: You know I haven't.
DALE: Then don't judge me.

Mmm... lung tissue

Today was another day of DNA isolation from lung tissue. I am teaching the Big Blue Assay to two people so they can do it after I am long gone! Today, I taught Bingxuan (a Chinese grad student that I worked on the a PCR project with and I like a great deal) how to do the isolation. Part of the process involves using a tissue grinder to break down the tissue and then lyse the cells. And today I broke the tissue grinder while I was grinding some lung tissue. So as the homogenate is spilling all over my gloves and the paper towels, I attempted to calmly open the package with the cell strainer (Bingxuan had gone to get the next tissue to grind) and then rinse the cells off my gloves with lysis buffer and scrape some of the tissue off the paper towel. As a result I can't imagine that I will get good DNA from the tissue but on the bright side the lung lobe from the rat was pretty large so I will be able to do a second isolation. And for the first time in about two weeks, I will be leaving before 4 pm (but coming in the weekend, so it really evens out) to watch some Hetty Wainthropp.

"Pittman, firing people can give you a pretty good buzz, but it's a poor substitute for killing. I realize that now." - Dale

Need some motivation?

See more Star Trek posters by clicking here

And more cat related poster by clicking here

"I don't know, Hank. Cotton working as a cop? He has a terrible human rights record." - Peggy Hill

Let's tune out by turning on the radio

After spending lots of time in the lab in the last few days, I was looking forward to unwinding a little bit tonight. I was really in the mood to watching the movie "Strictly Ballroom". Unfortunately, I sent the movie over to Rachel in Germany last July. So I did the next best thing, which was to download the soundtrack. Almost as good as watching the movie.

"To be honest, we're more into the invasion of privacy business, not the creation of privacy business." - Dale Gribble

Transpacking all day long

I am finally going to talk about the next step in the Big Blue assay which is called the Transpack reaction. The basic idea is to "package" the DNA into to a phage (a virus that infects bacteria) to infect the E. coli and insert the DNA into the E. coli DNA; the phage will kill the E. coli within a few growth cycle. Any mutation in the rat DNA is then put into to the E. coli and so I can identify the mutant. The E. coli is grown on a huge plate and creates a lawn of bacteria on the agar. Any E. coli that has been infected with the phage with die creating a clear spot on the lawn of bacteria. Mutant are identified by a clear spot with a ring a blue (basically the E. coli can breakdown the X-gal due the mutation for a few growth cycles and then dies). The basics of the Transpack reaction is this: I add the isolated DNA to a red tube (this is how what it is called in the manual, you have to buy the tubes from Stratagene) and then incubate it for 90 minutes. Then I add some of solution for a blue tube (again the official name in the manual) and then incubate it for 90 minutes. I forgot to mention that there are two different agar that I use a top agar and a bottom agar (the composition varies slightly between the two). Before I start the Transpack reaction, I make petri dishes that have bottom agar in them. After the second incubation is over I infect diluted E. coli with the Transpack reaction and incubate the mix for 15 minutes in a 5ml tube. Once the 15 minutes are up, I add 3-4 ml of cooled top agar to the mix, pour it on top on the bottom agar in the petri dishes, invert it and incubate it over night. The next day, I count the number of plaques (clear spots) and that determines the efficiency of the transpack reaction and also the number of big plates I need to pour to identify the mutants.


"Yeah, yeah, everything is one way then it's the other." -Hank Hill

Nobody puts baby in the corner

I saw an ad for Dirty Dancing and the fact that it will be shown in theaters in May (for 2 days) to celebrate the 20th anniversary. I'm not sure why it is being shown in theaters again. It's not like the viewing of the movie is enhanced by showing it on the big screen (and let's all admit the best thing about that movie is the soundtrack minus the weird Patrick Swayze song.) Maybe Patrick Swayze it trying to restart his career by reminding the American public that there was a five year period where he was on fire.

"I hear what you are saying. And I believe you believe it's important." - Hank Hill

Completion!

Here are some picture of my completed quilt top without any borders.












And here it is with the borders added:


"I have a new relaxation tape of me making ocean sounds." - Yogi Victor

I wish it was a musical number

So two of my sisters have asked for an explanation of the big blue assay that I am working on. I've decided to break it up into 3 parts. This assay is used to determine genetic changes in transgenic rats (you can also use mice or cells) following exposure to a substance. The rats have the LacI and LacZ gene incorporated on the 4th chromosomes which allows for you to determine genetic changes. The LacI is a genetic region that is control the ability of a cell to breakdown lactose, its acts as a repressor. If there is a change sequence in the area of the LacI (a region of about 1300 bp in length) it will generate beta-galactosidase. Beta-galactosidase will breakdown a substrate called X-gal which turns blue. And this is why it is called the Big Blue Assay. Although the company Stratagene has also created the same assay using a temperature sensitive based mutations but still gets called the big blue assay. The tissue that I am using is from a 28- repeat dose exposure study using PCB 3 and a PCB metabolite. I am using the lung from the male exposure for my particular experiment although last year I was helping Jim do the big blue using the liver from female exposure group. So the first thing that I do is isolate the DNA from the tissue. This involves pulverizing the tissue then adding enzymes that will digest the proteins, lipids, and RNA. Then I apply the DNA to a dialysis cup that will allow for small molecules to be separated from the DNA (the dialysis cup sits on the surface of buffer). The DNA will dialyze for about 48 hours and as a result will be very viscous. And then I use this DNA for the next step.

"How embarrassing! I thought I had removed the laces from this shoes." - Bill Dauterive

past and pending

Here are the other two blocks from my quilt:











BOBBY (shopping for a wallet): This one's pretty snazzy, and it looks like it could hold a ton of money. That way, if I wanted to impress someone, I could pull out a really big wad of cash -- bam! Dad, gimme a big wad of cash!
HANK: What are you trying to do, get us mugged?

one by one all day long ....

Here is one block (that is not completely pieced yet) of the quilt I am working on.



HANK: What kind of sick bastard runs a water-pipe through a stud without installing a nail-guard?
BOBBY: I don't know!