Um, thanks.

Re-reading my previous post (I do that to check that it reads okay, even after publishing. Maybe I'm narcissistic or something), I want to talk about something.

Google Searches.

I don't mind people finding my site through them, and if they read some of my ramblings before heading off to better written blogs, fine. But do you know how they get here? Half the time, they get here on searches which have to do with
(1) PhDs - are they worth it?
(2) stressed postdocs
(3) my boss sucks
(4) being a scientist sucks

(and say, 20% of the time on actual scientific topics, and 30% on random scientific stuff like barrycidal)

They somehow all radiate negative ideas about a being a scientist. Maybe my blog radiates scientific hate. I'm not sure how many of these people revisits my site.


Oh, and also....I have sorted out the outdated links on the right hand side. And on a side note, there is a reason for having baseball blogs at the top of the list for interesting blogs. All science and no play makes for a boring human being.

Google searches

So more stuff to add to the "searches" tag.. I've been slacking lately, but bear with me. I'm a busy person!

Query: Why you shouldn't do a Ph.D.
A: Hell, if you are Googling such a question, don't do it. It ain't worth your time.

Q: Ph.D. not worth shit
A: ......well, I'm glad I'm the one with a Ph.D. I've been there, and done that already. Don't want to do it again, mind.

Q: Chemical Biology trained as a chemist still not happy
A: Change your career. Do something different. Volunteer for something. Go to the gym. Get a life.

Q: Is it worth changing careers after 30
A: I believe so. If you are unhappy and determined to change your career, why not. The only person who will look out for you in your life, is you. Make life better for yourself.

But by Jeezuz, is there anyone who is happy out there anymore?!

Q: monologues on cricket
A: ......I still wonder how you got to my blog with that.

Q: Do I have to linearize my plasmid before stable transfection
A: At last, a scientific query! I was told by the Marlin that it is better to linearize it before stable transfection. And it makes sense - just think about it. For the plasmid to be integrated into the host genome, it has to be linear. If you put in a circular plasmid, it is up to chance where the cut is going to be. So, rather than risking the cells cutting your plasmid in between the insert (gene of interest), you might as well do that yourself and save your worries!

Q: Barrycidal AND clean an incubator
A: I swear by it. I recommend it. It's good stuff, and it keeps fungus at bay. Did I ever tell you that one about the "jellyfish" growing in the incubator? That was bad.

Q: Great scientists life story
A: Oh, thank you for the compliment!!!!

Google searches mysteries part deux

Okay, I've had some more Google searchers. Some of them funny, some of them...weird.

Q: day in the life of a bioinformatics scientist
A: I, presonally, don't know. I assume they are sitting in front of the computer most of the time, pretending to work when they are actually watching a YouTube clip, ogling porn, or masturbating over statistics.

Q: what's it like to be a scientist
A: Great for people not a scientist, shit for people who are.

Q: scientists in lab coats
A: Sexy, ay. I like my 5 year old lab coat with loads of ...what I think are developer stains on it.

Q: Weaver watch
A: Ahhhh, yes. Baseball. There was once a pitcher called Jeff Weaver; he once played for NY Yankees, I think he later went on to be a Dodger, and I don't know where he is now. Anyway, he was so so so terrible as a starting pitcher during his tenure with the Yankees, that the members of Channel 5's MLB show decided to follow his starts. And they called it "Weaver Watch". I believe he is the only pitcher who had a weekly update about their starts, on that show.

Q: why don't scientists speak English anymore
A: I thought we did. I wonder what this searcher was looking for.

Q: how to get rid of mycoplasma tissue culture
A: You can, theoretically, but if I had a culture contaminated with myco, I would bin it if I have a separate stock already available. I think there is an anti-mycoplasma thing, but I can't remember what it is.

Q: plan for working up a western gel
A: Ask your postdoc.

Q: FuGENE siRNA
A: I wouldn't. I have had bad experience with using FuGENE to transfect siRNAs. As, like, they don't. I would recommend a transfecting reagent made especially for siRNA, such as X-treme GENE (Roche), Oligofectamine (Invitrogen - and I have used it), DharmaFect (Dharmacon) etc. BTW, is Dharmacon related to the Dharma initiative???

Q: lipofectamine why wait 20 minutes
A: I think you need to read Maniatis. If you don't know why, you shouldn't be doing a Ph.D., and why do you think lipofectamine gets its name from?

Q: determine centrifuge time
A: Depends on the g required.

Q: dephosphorylation of vectors using CIP
A: I have done it, and I would suggest it. I just bought CIP from Pierce or somewhere, dephosphorylated the vector, and when you come to ligate the vector, you get far less false positives. As for why, well, do another Google search and go somewhere else. I can explain it but my fingers hurt...

More later. Definitely...

How do people get here?

I keep stats on this blog, and sometimes I get hits from people who put in questions in Google. Now, I can't say much on "The Ashes" or "How to make real butterscotch sauce", but I can answer some of them - or try anyway. So here are some examples which I recovered from my log stack:

Question: "why am I not getting any signal on my western blots"
Answer: There are several possibilities, ranging from the obvious, pathetic, funny, to serious. The obvious and quickest solution (if you are certain there are proteins on the membrane) would be to use a stronger ECL reagent. I find Amersham's - or GE Healthcare now - Advance ECL reagent is the best ever. Better than Pierce's femto reagent (I seem to get less background for the Advance reagent).
If that fails, do another western using more lysate/more primary antibody. I've never tried fiddling with the secondary, since it's the secondary (think about it). The more ridiculour reasons may be that you put the transfer cassette in the right way? I have made that mistake, and I have seen countless others make that mistake too.
Personally, I always check the transfer to the membrane is okay by doing a Ponceau stain. It takes 10 minutes, and will save heartbreak at the end of the experiment...

Question: "restriction enzymes broken freezer"
Answer: Now, I am guessing that that was someone who wanted to know whether restriction enzymes which were kept in a freezer, which broke down over the weekend, is okay to use or not. My answer is, don't bother and buy a new one. Most restriction enzymes are cheap, and if you find that your freezer has thoroughly defrosted, there is no point in even doing a simple restriction digest to see if it is okay.

Question: "beta mercaptoethanol what does it do?"
Answer: I always thought it was a smelly alternative reducing agent. Is it more stable than DTT (dithio-threitol)? I don't know. But I always put b-ME (as I call it) in my loading buffer. Alternatively, I have heard that you use it tissue culture, and that you use it on the bugs before transformation. I don't know why you use it in transformations.


I might do this more often...it's kinda fun...