Thermo Fisher Scientific field application scientist Dr. John Pfeifer explains the fundamental differences between PCR, real-time PCR, and digital PCR, including when each technology is most useful. He also shares practical guidance for designing, optimizing, and validating multiplex qPCR assays while improving throughput, sample efficiency, precision, and quality control.
PCR gets amplified, quantified, partitioned, and multiplexed in this practical tour through three decades of molecular biology innovation.
Dr. John Pfeifer, senior real-time PCR field application scientist at Thermo Fisher Scientific, joins Jordan and Lisa to explain how conventional PCR, qPCR, and digital PCR generate and interpret results differently. John describes real-time PCR as a versatile “Swiss army knife,” while digital PCR offers particular advantages for detecting rare targets against a high background. He then explores multiplexing, including how TaqMan assays use distinct fluorescent dyes to detect multiple targets in one reaction. The conversation covers the benefits of multiplexing for throughput, sample conservation, precision, and quality control, along with key considerations such as target abundance, Taq polymerase saturation, primer limitation, dye selection, mixed standard curves, and wet-lab validation.
In Career Corner, John traces his scientific curiosity from childhood books and theoretical physics to genetics, molecular virology, and PCR. He also recalls a demonstration gone wrong, a surprisingly restorative qPCR presentation involving Charles the goat, and the sales-rep conversation that changed his career.
Jordan Ruggieri 00:00
I have to admit on record here, I have not read a single word of any of these scripts. So, here we go.
Lisa Crawford 00:06
I did it about four minutes before the call, Jordan, so we're both crushing it.
Jordan Ruggieri 00:12
Why do I have a feeling that Matthew's gonna find that clip and put it in an episode at one point?
Lisa Crawford 00:16
We did zero work for this.
Jordan Ruggieri 00:17
Okay, here we go.
Jordan Ruggieri 00:22
Welcome to Absolute Gene-ius, a podcast series from Thermo Fisher Scientific. I'm Jordan Ruggieri,
Lisa Crawford 00:39
And I'm Lisa Crawford. Today we're delighted to welcome our colleague and resident qPCR genius, Dr. John Pfeifer, to the show. John is a senior real-time PCR field application scientist here at Thermo Fisher Scientific. Today he walks us through the fundamentals of PCR, qPCR, and digital PCR, how they differ, and why each technology is relevant in today's labs.
Jordan Ruggieri 01:01
You might say he amplifies our understanding of these techniques.
Jordan Ruggieri 01:06
You might. Regardless, it's a great conversation with someone who has been working with qPCR since it came on the scene. We hope you appreciate John's history and perspective as much as we do.
Jordan Ruggieri 01:21
John, thank you so much for joining us on today's episode of Absolute Gene-ius. We are thrilled to have you, and I know, knowing you personally, I have to say I am a little bit intimidated by how smart you are. So, I cannot wait to talk to you about science and multiplexing. So, thank you so much for joining us. Can you start by telling us a little bit about yourself and your role at Thermo Fisher?
John Pfeifer, PhD 01:46
Sure, I'm what's called a field application scientist for real-time PCR. I'm based in, or near Houston, Texas. I joined Perkin Elmer in 1991. This was before real time existed, so I was supporting PCR. And then in 1996 Perkin Elmer started selling the world's first real-time PCR system, and I thought to myself, “This is so cool, I want to support this technology.” So I've been supporting real time ever since.
Jordan Ruggieri 02:19
That is incredible, and what a journey. I can, I can only imagine the things that you've experienced with that rapid rise of qPCR. I can't wait to dig into that. But before we dig into that, I want to dive a little bit into some science here and talk a little bit about qPCR, digital PCR, what they are. Maybe for listeners who are a little newer to this, can you give a quick overview? What, what are what's what is qPCR? What is digital PCR, or PCR altogether?
John Pfeifer, PhD 02:53
So PCR began with the primary detection method as gel electrophoresis. And early on in PCR, it was pretty clear that researchers had a strong interest in gene quantification. One method involved was called competitors. But another method, which was published in 1992 is digital PCR. The idea there is to dilute the sample to the point where ideally you'd be dispensing, say, about one molecule of target per aliquot. Back in the 1800s, there was a French mathematician named Simeon Poisson, and he came up with an equation, not for digital PCR mind you, but for anything that follows that pattern of low probability events. An analogy that I like to use is imagine you have a cardboard box, and the bottom of the box there are 100 holes. Now imagine taking 100 rubber balls and just throwing them in the box. So we would not expect that there would be one ball passing through each hole. It doesn't work that way. Instead, some holes get no balls, some holes get two balls, three balls, and so forth. And so Poisson's equation then predicts those frequencies. So, going back to digital PCR, the idea there is you would calculate the percentage of non-amplifying wells, so those wells clearly did not receive target. We can, in a sense, reverse the Poisson equation to then deduce what must have been the original target concentration in the sample that was applied to digital PCR. Then of course 1996 Perkin Elmer introduced the world's first real time PCR instrument, and it was, the acceptance of that technology was a little tentative at first because the machine was like $100,000, so I can understand people being reluctant initially, but it quickly gained popularity. It was so much easier to generate a quantitative assay using real-time PCR, so that real time just grew and grew and grew. So then, in the early 2000s there was a company called Fluidigm. Fluidigm had developed a microfluidic system, so you could introduce assays and reagents and samples to create PCR reactions. Fluidigm was popular for a bit, but then I think they were trying to figure out, “What more can we do with this technology?” So in 2006 they developed digital PCR using microfluidics. Other companies started coming up with digital systems as well. So, with digital PCR, it's an endpoint detection. So, the question is, did a well amplify or not? And once again, if you calculate the percent of wells that didn't amplify, you can use that reversed Poisson equation to figure out how many copies were there. But in real time is a different story. There, we're collecting what's called the exponential data for real-time PCR. This is the early cycles in PCR, where there's plenty of Taq polymerase, primer, dNTPs. Everything's in excess, so that supports a consistent exponential-phase efficiency. So, as long as that exponential efficiency is continuing, we've established a quantitative relationship between how many molecules of the DNA target you started with, and when you start to detect the signal emerging. And so based on that we can then deduce how much of the target was there. Real-time PCR is, in my view, it's kind of like a Swiss Army knife. It's so versatile, you could do so many things with it. A digital PCR is more specialized, I would say. Any application where you're trying to take something really, really rare with a heavy background, like homologs, things like that, digital PCR might give you an advantage in that case.
Jordan Ruggieri 07:18
Are these technologies complement to each other, or do you see them more kind of competing with each other? And just kind of elaborating on, on, you know, is dPCR souped up qPCR when it comes to quantification, right? And are you going to, you get, you know, a little bit more, you know, for rare, rare events, rare targets? You get a little bit better of precision in that quantification, and so they play off of each other? Can you comment on some of that?
John Pfeifer, PhD 07:47
I think it would be closer to the truth to say that digital PCR, real-time PCR, are complementary. Again, real-time PCR is like the Swiss Army knife, you do all sorts of things, but there are certain applications, for example, mutation detection. Let's say you have a cancer allele that's present in a biopsy. Say someone biopsied some tissue, and let's say that the majority of the tissue that was biopsied is non-cancerous. There’s only a tiny percent that is cancerous. But what if you want to go further, then increase the sensitivity further? That's where digital PCR can come in. There if you dilute the DNA sample properly, you can boost its sensitivity by maybe tenfold, and that could be advantageous.
Jordan Ruggieri 08:53
Makes a lot of sense. Now, one element I wanted to dive into is a little bit around multiplexing. Can you give a little bit of background? What is multiplexing? High level, what is it, and why does it matter?
John Pfeifer, PhD 08:49
Multiplexing means, in real-time PCR that we're amplifying and detecting multiple gene targets at the same time. This would, of course, mean we're doing TaqMan, because it's very difficult to do multiplexing with SYBR Green, since SYBR Green is only one dye, it's one, one fluorescence. But with TaqMan we can choose different fluorescent dyes for each target, and our real-time instruments can discriminate those different dye signals. So that's a key factor of being able to multiplex. We do also offer what are called arrays. So arrays you could have 96 targets, 384 targets. But those are typically single plex, so there's only one assay per well, typically. But some people have referred to that as multiplexing, but I don't really think of it that way. But as far as why would you multiplex, multiplex in certain scenarios, can offer huge advantages. For example, if I had one target and one control gene for each sample I'm having to run the number of samples times the replicates, times the number of genes that I'm doing, that could end up being a lot of wells. But if I can do all those reactions in one well, so instead of target normalizer or target and control in separate wells, it's in the same well, I've boosted my throughput; I've reduced the amount of sample and reagent I'm consuming. I'm also improving precision, if that's part of what you're trying to do, and also you're improving quality control. Because you can imagine if you're a lab and you're trying to detect, let's say, a virus. If it's single plex, the technician could accidentally forget to dispense the right amount of sample for, let's say, the target versus the control, but you can't do that in a multiplex, because that same sample aliquot is being used for both the target and the control. So you've boosted your quality control, which is always a great thing.
Jordan Ruggieri 10:52
That makes a lot of sense. So, I mean, I like, I like your analogy. It's you might have thousands of wells, and in each one of those, you're detecting one target, but this is truly in one well, you're trying to detect multiple things within that well using different fluoresce versus the same, the same dye, the same fluorescent dye. One of the areas that that I find really interesting and intriguing about multiplexing is that concept of reducing the sample size. Can you elaborate a little bit on the benefit of that?
John Pfeifer, PhD 11:30
Yeah, part of it is utilizing less sample to get the same amount of data. If it's singleplex, you only get essentially one data point per well. Multiplex, you get maybe 2,3,4,5, who knows. So you're using, utilizing less sample to get that same amount of data. Also, it's multiplexing could be really useful if you have very limited sample. Let's say there's a target, it's very low in its abundance, and if you split that sample up, like in array, as an example, you split that up, then you could be that the target molecule that you wish to amplify, it didn't reach the well that contains that target assay, and so you missed it. But if you can put everything in one well and you're not missing anything.
Lisa Crawford 12:20
Why wouldn't somebody use multiplexing? It sounds like it's an advancement that, like, everyone should use in the lab, or is it only for specific circumstances?
John Pfeifer, PhD 12:28
There are different scenarios. There's a multitude of scenarios in real-time PCR. So, if you have a scenario with a small number of targets, then multiplexing could be really advantageous. The easiest, simplest example is you have one target, one control assay. And if I was going to run, let's say, ten samples in triplicate. In singleplex, that would require 60 wells. But if I were to duplex that, I could only, I could run the same experiment, get the same data with only 30 wells. So I've just saved half on my reagent, half on my sample, half on my thumb, because I'm not pipetting as much, which is nice. And then there's the precision benefit and quality control benefit as well. But let's say I've got 48 targets. Well, there's no real time machine in the world that can differentiate 40 different dyes. It just doesn't exist. You have to think, what's the better fit here? I could try to generate multiple multiplexes for 48 targets. That's going to be kind of a brain buster, I think, or I just get an array. Even just a 96 well, our simplest array TaqMan array plate. 96 wells, you could run that easily as an array. You order it, here it comes, and you didn't have to worry about multiplexing, as far as the decisions, which dyes do I choose, which essays do I primer limit. I have to submit the assays to our multiplex support tool, etc. etc.. All that goes away. You go right into an array situation.
Jordan Ruggieri 14:08
I think the other end of this is, how complicated is it to set up a multiplexing reaction? I mean I can assume, you know, I remember my time in the lab, spending weeks, sometimes trying to figure out one assay, right? In terms of primers and dyes, and “Why is it not working,” and “What's a problem here?”, and “How do I validate and verify? and all that kind of fun stuff. Do you have to do that 3,4,5, times if you're, if you're multiplexing?
John Pfeifer, PhD 14:34
Good news here is that in the late 1990s when again PerkinElmer, this was the Applied Biosystems division of PerkinElmer, created real-time PCR, that they quickly understood the complexities of PCR. So like what you're saying when researchers were first working up their primers and running the gel the first time, sometimes the results didn't look very good, and then they would embark on what's called optimization, maybe optimizing the primer concentration, the annealing temperature, you know, all sorts of things, and so that's when our researchers said, “Well, we've got to, we got to fix this problem, we can't be dragging a boat anchor with real-time PCR.” So that's when they embarked on the creation of a streamlined system for real-time PCR. So everything's universal. Universal primer and probe design rules. Universal primer and probe concentrations. Universal magnesium, universal cycling, and so forth. So that was a huge simplification. The multiplexing is kind of a building from there, that's the foundation, I think. Our universal system, so universal probe concentration, you never have to change that, you never optimize the probe concentration. Universal cycling, well, obviously that's beautiful for multiplexing, because everything in that well has to amplify under the same cycling conditions. There are a couple issues to be aware of. One of them is called Taq polymerase saturation. So, saturation means that if we were to use our universal singleplex primer concentration in a multiplex, if the target in that sample was really abundant, much more than other targets, then what we run the risk that the amplification will continue beyond exponential and well into linear phase, which is caused by a saturation of the Taq polymerase. So just so much of that amplicon, it just the polymerase can't do all the work, and that would starve the amplification of lesser abundant targets in the same multiplex. The solution, though, is actually pretty simple. We just order the assay primer limited, and that prevents that saturation from happening. And you can order any of our assays that way, either off the website directly or using part of our company called Specialty Oligos, and they'll be happy to make them primer limited, so the research doesn't have to lift a finger.
Jordan Ruggieri 17:06
There seems to be a lot of considerations, right, in terms of multiplexing. Can you talk a little bit about, you know, maybe what are some ways to get started if somebody is looking to multiplex or has multiple targets? Maybe even from just a simple, “Hey, check out a TaqMan assay, right, a pre-designed assay,” right? Like, how would you recommend somebody getting started, they want to multiplex, they want to look at multiple targets, what are kind of some steps you would recommend?
John Pfeifer, PhD 17:36
Sure. Step one, of course, you need to identify well, what are the targets of interest. We do have the custom TaqMan assay design tool. If a researcher doesn't have existing primer probe sequences, the tool could design assays for them. Researchers will often obtain primer probe sequences from the literature. So that'd be step one, identify your targets, and then figure out where they need to come from. Could they come from us, Therma Fisher, or other sources? Then I would suggest going to the multiplex support tool in the assay design web page, and submit them. So you can either submit an assay ID number or you could submit primer probe sequences, whatever, whichever you prefer to do, and then they will do this in silico check to see if there are potential adverse primer probe interactions. Let's assume that none of the targets are predicted to have interaction problems. So the next question would be, what do you want to assign them for the reporter dyes? There has been a misconception out there that it's better to select dyes that are furthest apart in the EM spectrum, and that's really not required in our systems. Maybe there are other real-time vendors that that's a requirement for them. I don't know, but not in our systems. As long as the dyes are properly calibrated in the instrument, you can have the dyes adjacent to each other. In fact, when we first came out with real-time PCR, the two most common dyes used in multiplex were FAM and TET, and FAM and TET overlap a lot, but yet the system that we used to describe the dyes was very powerful and could separate the dyes very cleanly. Then think about which assays need be primer limited, if you're not sure about which assay could potentially be more abundant than the target, be more abundant than another target, the safest thing to do, just have all primer limited. Although there's something to consider if you're considering a one-step chemistry. So one-step is where you have a single reaction that'll perform both reverse transcription and then without user intervention, goes straight into PCR. The priming for cDNA synthesis in the one-step chemistry is accomplished by the reverse PCR primer. Therefore, the concentration of the reverse PCR primer can play a role in reverse transcription efficiency. My personal suggestion would be, if you're attempting a one-step multiplex, you may want to consider primer limiting only the forward primer and leave the reverse primer at universal. That should maximize reverse transcription efficiency for that RNA target. Once you've made all those decisions, order the assays. I'm going to suggest small scale, the smallest scale you can get, and I'm also going to suggest get them in the single tube format, that is, primers and probes mixed together in a single concentrate. And the reason I'm suggesting that is, multiple reasons, one is it's always useful to be able to create an assay master mix with a consistent primer probe primer concentration ratio. We want to keep those concentrations consistent, and a single tube assay is a way to do that. If, for example, you would be dispensing the primer working solution for the forward primer, primer working solution for the reverse primer, the probe working solution. Well, you see, there's a lot more pipetting involved. There's more potential for error, and this is particularly important in multiplex because it's important to have primer limitation, and I have examples with customers where they were hand pipetting primers and probes individually, and sometimes they get it wrong. The primer is not primer limited, they think it is, but it's not. And so, sometimes you get that Taq saturation effect I was talking about. You get a suppression of the lesser abundant target, which ruins the multiplex. With a single, with the small-scale individual tube assays, this gives you the flexibility to then do multiplex testing. Now, I mentioned before about submitting the assays to our multiplex support tool, and that's great. It's in silico analysis, but if you really want to be sure that the multiplex is going to perform as you would hope, is to do wet chemistry testing. This is the ultimate in quality control. And so by having them singleplex initially, the best way to test for multiplex performance, test each essay in pairs in what's called a mixed standard curve. And we have an example of that on one of the product web pages, where they did a multiplex with a one-step chemistry. So one target is held constant, the other target, you do a serial dilution, you mix them together, so you're trying all the different ratios of the two targets that potentially exist in actual samples, and you're looking for the maintenance of linearity. As long as everything stays linear, you're good. So, once you've done all that testing, now you have powerful data that says “This multiplex is ready to go. It has the performance that I need, that I expect.” Now, you can tell, especially all it goes to formulate all the assays together into a single tube, and your life is now so much simpler.
Jordan Ruggieri 23:28
So, John, I have, I have one more question. I want to hop into some of the career questions, and even talk further about your career towards the end, but is there a difference in multiplexing fundamentally between real-time PCR and digital PCR?
John Pfeifer, PhD 23:44
Well, multiplexing is not as emphasized, I think, in digital PCR, because again, there's a range of target concentrations that it needs to be for Poisson to take effect. For example, if you add too much target, you could saturate all those 20,000 wells. So in the Absolute Q and in the QuantStudio 3D, I'll just call them a chip, for sake of argument, is divided into 20,000 little, tiny wells. And so we want to have a substantial number of those wells not amplify in digital PCR, so that we can apply the reverse Poisson equation with statistical confidence, shall we say. So, if you have a multiplex where the two targets are always going to be very similar to each other in their abundance, that could work, you know, for multiplexing in digital, but if you have a situation where the targets can vary in their abundance significantly, then that's going to be more difficult.
Jordan Ruggieri 24:52
So it sounds like there are there are some differences in considerations when it comes to multiplexing, but in different situations you may choose one over the other, but they're both theoretically doable as long as you, you take into consideration some of the kind of standard rules for multiplexing?
John Pfeifer, PhD 25:11
Yeah, follow good procedures. More good news is that we have a wonderful technical support team, also there are FAS, field application scientists, who can guide the customer through these steps, step by step to make it as simpler and as easy as they can make it.
Jordan Ruggieri 25:33
For 30 years, qPCR has helped power discoveries in gene expression, oncology research, infectious disease studies, agricultural biotech, and so much more.
Lisa Crawford 25:43
And Thermo Fisher continues to build on that legacy with innovations designed to improve qPCR accuracy, workflow efficiency, and data confidence.
Jordan Ruggieri 25:52
To learn more about 30 years of qPCR innovation, visit thermofisher.com/qpcr 30. That's thermofisher.com/qpcr 30. Products are for research use only, not for use in diagnostic procedures.
Lisa Crawford 26:06
And now back to today's guest.
Jordan Ruggieri 26:10
Awesome. Well, I think that means it's time for Lisa's Career Corner. Lisa.
Lisa Crawford 26:16
Moving along away for a little bit away from the science, but yeah, what we like to do in Career Corner is just kind of learn a little bit about the person behind the science. So you know, tell us how you even became interested in science, not necessarily, you know, in college, but you know, as a child, was it something that you were particularly drawn to, and what kind of sparked your initial entry into the field?
John Pfeifer, PhD 26:37
It was the summer between my fourth and fifth grade that I was bored. Asked my mom, “What could we do?” She took me to library. I found a series of science books. They were called the Time Life series, and some were about the planets or the ocean or all kinds of really cool stuff. So I was just fascinated, absolutely fascinated. Read all the books, I said I want more books. So go to library and I got this book about Albert Einstein, the theory of relativity, and all that stuff. So then it was the beginning of my fifth year, and the teacher asked all of us to stand up one by one and say your name and what do you want to be when you grow up. So I stood up and I said I want to be a theoretical physicist. The teacher stood there with her mouth hanging open for a few moments. She said, “Do you know what that is?” I said “Yes. Oh, it's like Albert Einstein. I want to be like him.” “Okay, you can sit down now.” So, yeah, so but a key person that I really want to mention that led me into molecular biology was my high school biology teacher, Mr. Culpepper. He was, he was amazing. He was teaching molecular biology before I even think there was that as a terminology, and I just, that's what led, that's what inspired me was him being a teacher, teaching that material. It just, I don't know, went right into my brain like a sponge. At Cornell as an undergrad, I got my bachelor's in genetics. So then I, when I was in graduate school, I studied, you could say, either microbiology or molecular virology. So I began to develop an interest in viruses. There was a paper published when I was still at Cornell where they sequenced SV40, a Simian virus, and I was thinking about this could be really cool for like genetic engineering so I got really excited about the molecular basis of viruses. So that's what I worked on in graduate school. Now, and I continued on with this, was focusing on human respiratory virus, particularly like influenza virus. So that's when I went to Harvard as a postdoc and worked on making mutations in the viral nucleic acid and transfecting cells and looking at what happens when you do that.
Lisa Crawford 29:16
What was it that kind of led you from that more academic, you know, virology research into the more technical aspect? Because you talked about, you know, joining PerkinElmer and just really kind of falling in love with the technology? What was it about the technology that kind of drew you away from the more traditional, I guess you would say, academic research path?
John Pfeifer, PhD 29:36
I had begun to feel, even as a graduate student, an attraction towards industry. My sense was in academia everybody is like their own island. You kind of work alone and you're in competition with other people, and it didn't feel good to me. I wanted to be part of a team, a team with a unified goal. Everybody's working together to achieve these goals. And the opportunity, when I was at in Houston, I was working at Baylor College of Medicine, and this opportunity just appeared, and so I leapt at it. I thought, “This is going to be great.” And it was supporting PCR, this was before real time existed, but it was supporting PCR. The PCR is still fairly new at that point, and so I was explaining to researchers how to use PCR. I was doing seminars about PCR, training them, that sort of thing,
Lisa Crawford 30:40
Something we love to ask everyone, and bear with us here, but we like to ask everyone, what is your most embarrassing moment that you've had in the lab? And then maybe, what is your proudest moment that you've had?
John Pfeifer, PhD 30:53
Well, I think the most embarrassing was I was demonstrating the, this brand-new real-time machine again, real time was brand new. This is the very first model of real time instruments. It was called the 7700, incidentally. And the way it worked is you had this handle, you would rotate the handle, push the heated lid back, place the plate, put the heated lid forward, turn down the handle, and in that, that's how you could load the plate. So I was, I had a room full of scientists, I was demonstrating how you twist the handle, and as I did that, the entire handle assembly fell apart, so that was pretty embarrassing.
Lisa Crawford 31:38
Keep going, pretend, or could you not even open it after that?
John Pfeifer, PhD 31:42
So I tried to play it off as “Well, obviously, this, the screw wasn't screwed it all the way, it got loose. Should be pretty straightforward to fix this problem. No worries.”
Jordan Ruggieri 31:56
Couple rolls of duct tape later.
Lisa Crawford 31:58
Yeah, pay no attention to that part.
John Pfeifer, PhD 32:01
Oh boy.
Lisa Crawford 32:02
What would be a proud moment that you remember?
John Pfeifer, PhD 32:06
Let's see, what's a good one? Okay, this may be little off the wall, but I'm going to go for it. So I was doing a training. It's kind of an agricultural type college. It was probably January, winter, cold. And I go into this lab to train them, and they bring in a baby goat. And they explained that a mama goat, they study goats, that's what they do. Mama goat had given birth to two babies the previous night. One baby was smart and spent the night next to his mommy, stayed warm, no problem. The other goat was less intelligent and spent the night next to the metal wall of this shed, and it was hypothermic. So the goat was just lay, it was limp, had showed no signs of life other than, I guess, breathing, and they were not sure if the goat was going to make it. So, they start trying to feed it some warm milk, and it was time, so there was an initial period in the training where I showed the instrument. Then we go to the conference room for a presentation, so they decided to bring the goat with them to the conference room. So while I'm presenting they're still feeding the goat and massaging it and things, and the goat starts getting better during my presentation. And by the end of my presentation, the goat was so good that it followed us, it actually could walk on its own and followed us back to the laboratory. So my thinking is “Real time PCR, my presentation was so good, it saved a goat.”
Jordan Ruggieri 33:55
Oh, that's the best story I think I've ever heard.
Lisa Crawford 33:59
That goat is now tenure track at Texas A&M.
John Pfeifer, PhD 34:04
And they named the goat Charles, by the way.
Lisa Crawford 34:06
Charles the Goat, I love that.
John Pfeifer, PhD 34:10
I've got pictures of Charles,
Jordan Ruggieri 34:10
Your presentation was so lively, that the livestock jumped up. John.
John Pfeifer, PhD 34:19
Yeah, that's what I'm gonna go with.
Lisa Crawford 34:23
Herds love science, this is well known.
Jordan Ruggieri 34:26
Oh, that's the best one. I, that takes the cake, John. That was wonderful.
Lisa Crawford 34:34
I just have one more question for you in this track, really, and that would be, what advice would you give to younger scientists who are maybe just starting out in their careers? And you know, something that maybe you wish you had known, or that you look back on and would like to share?
John Pfeifer, PhD 34:50
There's two things that I guess I would talk to a young, say, a young researcher about. One is consider quality control. So, quality control, I think, is a really important part of every human endeavor, but particularly in science. Either creating a set of experiments, running scientific tests, what have you. That will help ensure that you're producing accurate data, which I think is one of the top goals of any scientist. Also, if you learn about quality control and you embrace the concepts, quality control, quality assurance that will help open doors, I think, for other opportunities. For example, in basic research, there's no real rules you have to follow. There are no regulations you have to follow. So, maybe quality control isn’t important there. But if you move outside, you move into industry, move into diagnostics, quality control there matters a lot. So, I think that would help open the door to more opportunities. And the other thing is, be nice to sales reps. I mean, that's, that's how I got this job. So there were many scientists, colleagues that viewed science, that viewed sales reps as an annoyance. “They're just trying to sell me something I don't need it,” and you know that sort of thing. “They don't know anything about science. Why should I talk to them?” You know that kind of thing. So very negative attitudes. My view, though, was that sales reps are trying to help you ultimately. Mainly they're trying to fit one of their products in their portfolio to your work, but that's a plus, because you wouldn't necessarily know about those products until they tell you about them and those technologies might be great for your scientific research. So be nice, and that's how I got this job. I talked to a sales rep, we had a great conversation. I said, "Hey, let's go grab lunch." We had lunch together. Great conversation. I leave. She gets a call saying, "Hey, there's this job position open, we need someone who's both technical and has a personality. Do you know anybody?” That's how I got this job.
Jordan Ruggieri 37:22
John, thank you so much for being on today's episode of Absolute Gene-ius. It was thrilling. I learned a ton. Really enjoyed the conversation. Thank you so much for your time.
John Pfeifer, PhD 37:31
Well, thank you. I appreciate the opportunity.
Jordan Ruggieri 37:34
That was Dr. John Pfeiffer, senior real-time PCR field application scientist at Thermo Fisher Scientific. We've got more great conversations and Science Snapshots coming soon. In the meantime, stay curious, and don't forget to subscribe wherever you get your podcasts, so that you don't miss a single episode. This episode of Absolute Gene-ius was produced by Sarah Briganti, Matt Ferris, and Matthew Stock. All products mentioned in this episode are for research use only, not for use in diagnostic procedures.
Lisa Crawford 38:02
Look at you, off the cuff like a pearl.
Jordan Ruggieri 38:06
It's not off the cuff. If you, if you knew my background of that little line right there, you have one too it should be tattooed on my forehead.