With advancing years, I find myself taking on the annoying traits of the older people I used to secretly pity. Yes, the young really don’t know how good they have it, and no, the world would not actually be a better place if we all spent every waking hour wearing tie-dyed T-shirts, protesting about injustice and trying to save the whales.
But the opposite seems to be happening to me in my scientific life. When I was just starting out in research, I was fascinated by the rise of kits – those experiment-by-number packs which must surely represent the ultimate lure of scientific youth. The lab in which I did my Ph.D. research wasn’t prone to frivolous spending, but I would watch colleagues in richer labs with envy. These kits never opened doors to techniques that would otherwise not be possible: they just made things easier. It was the same reactions, packaged up in neatly nested bottles and tubes with a handy instruction manual. Typically, the kits involved some pretty major short-cuts, were often less toxic and, probably most admirably, freed you from the tedium of having make all the solutions yourself. Scandalously expensive, they represented all that our betters were constantly warning us against: mental laxness, fiscal extravagance and (the unuttered subtext) the worst possible species of moral weakness. What sort of real scientist (we were told) would spend $300 on a kit when he could cobble together the necessary chemistry for a fiver?
Urban myths started to circulate, too, near the end of my Ph.D., about candidates caught out in their oral exams when being asked to explain exactly what Solution A did to Substrate B in that shiny blue box. Today’s youth, we heard, had become mindless drones, performing manipulations without truly understanding what they were doing. All of science would surely shudder to a halt.
But, you know what? That never happened. The kits persisted, and increasing numbers of labs started to use them. Time is also money, and what is a better use of time, spending two hours fiddling with an analytical balance and a pH meter, or doing a real experiment with a reagent you prepped in half the time using a kit? I certainly don’t miss the toxicity: as someone whose trendy 1980’s jeans are probably still quarantined in a radioactive landfill somewhere near Bethesda, and who once spent a memorable couple of hours in a Seattle ER getting phenol rinsed from her eyes, I appreciate the clean, safe workflow of modern luminescent detection systems and nucleic acid-binding columns. And as someone who once wasted four months of her life because a technician inadvertently messed up the recipe for a simple buffer, I like the reproducibility and reliability of the pre-made solutions. I still do a lot of stuff the old-fashioned way, but I’m increasingly succumbing to the allure of ease that kits provide. I know they are more expensive, but I do wonder how much money they save in the long run.
At the moment, I’m elbow deep in a wondrous new kit. After successfully completing the three constructs I promised to make for a colleague, I’ve got a week left to recombine them into insect virus ‘bacmids’. I may have a Ph.D. in retrovirology, but I don’t know my baculoviruses from my backside. With the kit, I don’t have to. Yes, I’m not ashamed to admit that I’m just following the instructions line by line without having the slightest idea of the biology that will transform my miniprep DNA into one of these do-hickeys capable of creating transgenic insect virus particles. If it all goes to plan, it doesn’t matter. (I feel deliciously naughty just admitting that in public.)
So that’s me: the aging kit convert. I think I’m just too old to do molecular biology in a cardboard box in the middle of the road like my grandma used to. Is that so wrong?
I think ‘lack of moral fibre’ is what we used to call it.
Now, I know I have been Mr Grumpy when it comes to kits, but I actually think some of them have their place. And I was in the industry, once. Hmm, I might have to blog about this myself, as I’ve been promising it for a while anyway.
So you don’t fashion all of your plasmids out of twigs, then?
Plasmids, no.
Proteins, however, I make from the freshly distilled nectar of the Oomagoolie Tree.
Also known as Buffer 3.
HA! I heard the same urban legends. I wonder how many people actually got that kind of question? The stories certainly inspired me to do some pretty intensive cramming in the week before my viva, anyway… especially after my PI told me to make sure I knew what every single ingredient in every single experiment did!
Phenol in the eyes sounds awful. I got a tiny spot on my skin once, and it raised quite an impressive blister.
They basically slap what look like very thick contact lenses to your eyes, connected via tubing to saline drips. They drip about 4 liters over your eyeballs over the course of an hour. Of course your eyes are open through all of this — seriously seasick the whole time. I was lucky that my lab was in a hospital building, so the ER was just a short walk away. I also benefited from a quick-thinking colleague who stuck my face under an eyebath station only a few seconds after the incident.
In my oral exam, I had to go to the board and draw out the entire process of reverse transcription. Not as east as it sounds!
I had to explain Dicer/Drosha/RISC etc. at my defense at my defense, even though I used RNAi as a tool, not as a field of research. But I can imagine that students won’t have to know that five years down the road, and knockdown will just be a “kit” in its own way.
Nobody asked me about LyseBlue.
Oh, and I even had to make a figure in my thesis showing the RISC pathway. But my thesis is CC licensed, so anyone can now use my figure. (And then, ideally, the credit would somehow count towards a magic new kind of index that says how awesome I was/am as a scientist. But that’s another topic of conversation.)
I don’t know my baculoviruses from my backside
Thank goodness you do all your research standing up, then.
Damn, Henry got to it before I did 🙂
I just experienced the reverse: A genotyping kit that would have saved tons of time, and even cost less than our standard protocol. Sadly, it gave horrible results. Well, at least now I have gel pics to show to students for any imaginable error…
Goodness, Eva. I don’t think I remember the exact specifics of the RNAi pathway now either, and I do it every day. But then, nobody ever quizzed me about the precise biochemistry of restriction digestion or ligation. It’s all about familiarity, I suppose.
I wish I knew how LyseBlue worked, because my buffer has stopped doing the blue thing after a few weeks in the fridge. That’s when I realized how much I like it.
It isn’t just kits either. I had incredulous looks from some of my colleagues yesterday when I said I could make competent cells.
@ Kate – your comment reminds me of the apocryphal story of a road sign which read
HARWICH FOR THE CONTINENT
beneath which some wag had written
FRINTON FOR THE INCONTINENT
Alexander, I haven’t had any bad experiences with kits yet, but that is the main drawback – when they don’t work, it’s hard to troubleshoot because you don’t always have enough information.
“pH meter”? pH meter??? Have the youth of today forgotten how to do titrations?
For Tom:
drip… drip… drip… DAMNIT!!!!!!
[repeat as necessary]
@Kate – me too=. And I shocked a number of people in these parts a few years ago by suggesting that they could resolve certain questions easily by running a Southern blot. That suggestion did not go over well, of course.
@Eva – on my reclass exam, some
jokerprofessor asked me if I could draw a Holliday junction, since my proposal talked about recombination. Iknowsuspect that this was a defensive tactic to hide the fact that he hadn’t read the thing, and thus had no sensible questions to ask.Heh. And some people don’t like NanoDrops…
Whenever I suggest to fly people that they might want to do an in-vivo labeling to see if their protein truly is phosphorylated, it seems to be akin to the suggestion that they mug their granny. But as far as I know there is still no reliable non-radioactive way of doing that. (I tried a number of reagents and antibodies back in the day, but none really worked as well as an atomic pile’s worth of 33P.)
Back in the ancient times when I was in grad school, one professor used to go off on a much-parodied rant about “you post-cloning kids”, and how spoiled we were because we could get EcoR1 from the freezer, rather than having to grow E. coli and isolate it ourselves. I have personally updated the rant to “you post-genome kids”, as my entire thesis project (which involved sequencing and comparing a 10kb stretch of regulatory regions) could now be done in a matter of weeks.
I felt the same way about spoiled children and kits (or pre-mixed solutions–you really need to buy PBS, you can’t just make it?), but now I think it’s time to move on. Now we should embrace kits, but rant at outsourcing, things like having a company make a transgenic mouse for you. Back in my day, we did our own microinjections, grumble, grumble…..
I suspect, David, that a lot of the reaction you describe is down to secret envy: why should young upstarts enjoy things that weren’t available in my day? My plan is to age gracefully and try to see change as a good thing (within science as well as without) – not always so easy!
@ Richard W
ooh interesting method, what sort of competence do you achieve and have you tried it with anything other than XLIBlue?
@Kate and Richard> Ahh… the times in the cold room with a box of ice and water making “slushy cold water” to chill the E.coli with…. And a huge E-flask to move around in the box 🙂
Jenny> I think there is something to the thought to know “what the kit is doing”, also called “knowing why you need to put solution A in B”. A student asked me about that when we went through the ELISA process and using HRPmarked antibodies. Let’s just say it was fun and I felt superold going with “when I did this when I did my PhD…”
If nothing else, the kits do make it a bit more consistent. (As long as the company don’t change their purification protocol of course…. and forget to tell you….)
OK, here’s one for the purists:
would you rather make a curry from the freshest of individually selected ingredients, or bow to Mr Patak?
(Just got back from Brick Lane. Wow)
Answer: don’t care, as long as the product produced is tasty enough for what is required subsequently. If it takes half the time and the chances of success are more reproducible, then by all means pass the reconstituted mango chutney packet.
Asa, for basic kits that are just making tools (like a miniprep kit), I honestly think that you only need to know how it works if it doesn’t work. If it does? Shrug.
Heh.
Problem there is that you don’t know in advance if it’s not going to work.
Actually, the biggest reason for knowing how kits work is that you can then make them up yourself, for a fraction of the cost, sometimes using exactly the same reagents as you would be sold in the kit.
Thereby wasting aforementioned time and risking error.
Having said that, most of the kits I use involve technology that I can’t readily replicate — for example, enzymes and vectors for advanced recombination cloning techniques. I’m happy to let the experts do it.
A “kit” really isn’t that different than ordering a restriction enzyme from, say, NEB, is it? Purified reagent, with co-packaged buffers and instructions.
Biggest problem with kits is people not reading the directions. Ask me about PCR cleanup kits prior to very expensive next-generation sequencing reactions sometime. But only if you have a large chunk of spare time that you don’t mind sacrificing.
@Kate – it was a while ago. We used those XL1Blue cells for standard subcloning as they were cheap and could be stored in the fridge (no time wasting thawing them out). I doubt they had high enough transformation efficiency for, say, making a cDNA library (does anyone do that any more?). But for regular subcloning they’re the business.
Never tried with other strains, although in principle I don’t really see why it shouldn’t work. I think the reason to use XL1Blue was that they’re Tet-resistant, so you can grow them under selection – although I don’t imagine this is really that important. I heard anecdotally that in some European countries you can’t get Tet-resistant XL1’s because, well, they’re Tet-resistant.
Be interested to know if this protocol works with other cells, if you feel like doing the experiment… hardest part of the protocol is pH’ing the KMES, honestly.
Oh, and maybe 10^7 to 10^8 per ug as I recall, definitely not 10^9 though.
I would be very happy if I could live safely in the knowledge that I never have to use a pH meter again as long as I live. I love the high-resolution litmus paper, though. So aesthetically satisfying, and somehow I believe it more.
p.s. Richard G probably used to isolate his own restriction enzymes.
Yes, and mouth-pipette 32-P, which might explain a few things.
I’m with you on the pH meter, Jenny. Evil things.
I think you misunderstand me, Jenny. I was thinking specifically of the Quikchange mutagenesis kit.
In it you get all these lovely boxed reagents, but you can buy exactly the same reagents, separately, and do exactly the same thing, at much less price. You can buy Pfu, DpnI and the nucleotides and buffer in one box; or separately. Which turns out cheaper. The same things! The identical protocol.
Similarly with the RT kits from Invitrogen. You can get the enzyme and the 5x buffer, or you can get a kit with umpty other reagents that you never use.
Ah! OK. That makes sense.
That reminds me, I was thinking of setting up a small business selling off the ten million blue tubes of pUC18 control in our -80…
Hey! I made sense!
Perhaps you could make necklaces from the plasmids?
I am now anxiously anticipating Labart to go with Lablit. You know you want to.
I second that.
Labartlabartlabartlabradorlabartlabartlabart.
A surprisingly disturbing juxaposition of letters this close to midnight.
LabLit already does art – various pieces about it, and actually a few works themselves. We’d love to have more.
Labartlabartlabartlabradorlabartlabartlabart
For some reason I am now thinking of large black dogs.
Plonkerplonkerplonkerplonkerplonkerplonkerplonkerplonkerplonkerplonker.
The caliber and intellectual content of this salon never ceases to amaze me.
We aim to please.