Good things come in little packages, is the theme for this episode. Join us to learn about micro RNA (miRNA) and how digital PCR is being used to understand its role is disease states.
Before the 1990s, small bits of RNA were considered junk by most, but the 1993 discovery of microRNA (miRNAs) began to reveal that bits of only 19-24 nucleotides of RNA can have an important gene regulation function in cells. Since their discovery, there has been a flurry of work to catalog known miRNAs and understand their functions, which include being tied to specific disease states such as leukemia.
According to our guest, Dr. Guy Novotny, Molecular Biologist at Herlev Hospital in Copenhagen, it’s now relatively easy to identify a miRNAs and follow their expression, but to figure out what they’re actually doing is a real challenge. We hear how he and his team have recently adopted digital PCR, and the benefits that come with it, to study microRNAs and figure out what proteins they’re regulating the expression of. This includes basic research, where Guy is “adding to the big pile of data that’s existing out there,” and he also does clinical research that has a closer connection to specific disease states and subject outcomes. As always, you’ll get to learn about his career journey and learn that there’s really not much that cake cannot fix.
Visit the Absolute Gene-ius page to learn more about the guests, the hosts, and the Applied Biosystems QuantStudio Absolute Q Digital PCR System.
Jordan Ruggieri 00:00
Man, I'm going to be singing on the next podcast episode. Oh, dang it.
Cassie McCreary 00:16
Welcome to Absolute Gene-ius, a podcast series from Thermo Fisher Scientific. I'm Cassie McCreary.
Jordan Ruggieri 00:21
And I'm Jordan Ruggieri. And today, we go big by going small with our micro-RNA Gene-ius Dr. Guy Novotny.
Cassie McCreary 00:29
Guy is a molecular biologist in the hematology and pathology departments at Herlev Hospital in Copenhagen, Denmark. He's worked in a variety of fields within microbiology over his career in science, from the research lab to the clinical setting. And outside of his incredible work, he's an avid sci fi reader with some book recommendations at the end of the episode. So, sit back, relax, pop in those micro sized headphones and enjoy.
Jordan Ruggieri 00:55
You know, when I was a kid, I was in you know, had to sing, at like a preschool. And one of the songs we had to sing was "Good things come and little packages, little packages. Yes, indeed."
Cassie McCreary 01:08
Wow what a banger. Jordan's going to top the radio charts right now.
Jordan Ruggieri 01:16
All right, Guy, thank you so much for joining us today on Absolute Gene-ius. We're very excited to have you. Would you be willing to give us just a brief background of yourself and some of the research that you conduct?
Guy Novotny, PhD 01:27
I'm a molecular biologist, PhD, and I've been working for many years as a researcher, often with micro-RNA and RNA as a subject. The last maybe a decade or so I've been working hospital centers, pathology centers and molecular diagnostic centers with a lot of hematological samples.
Cassie McCreary 01:52
I have a question, yes. For our listeners, for those who might not be familiar, what is micro-RNA?
Guy Novotny, PhD 01:59
So, micro-RNAs, when I was doing my master's degree of work, they weren't very well known. But they call them micro-RNA because RNA molecules are thousands of nucleotides long. Micro-RNAs are only around 20 nucleotides long. So, that's why they call the micro-RNAs because they're pretty small RNA molecules, even though as a molecule itself, RNAs is quite large, even a micro-RNAs. To start with, no one really knew what they did, but they actually find out that they regulate the expression or the translation of other messenger RNAs. So they are they're like a negative regulator of protein expression. And since they're so small, it's difficult to figure out exactly what they're regulating. So, it's pretty exciting how to discover all these new micro-RNAs, then you have to figure out what they're actually regulating.
Jordan Ruggieri 02:45
Interesting. And do they, by regulate, do they up regulate? Do they down regulate?
Guy Novotny, PhD 02:50
Most often, the most common method would be to down regulate or to prevent translation of a messenger RNA. So, you down regulate a protein level with the micro-RNAs. But there are, there are some articles, some researchers say that they do the opposite. They can upregulate as well.
Jordan Ruggieri 03:07
Very interesting. How do you identify specific micro-RNAs?
Guy Novotny, PhD 03:11
Initially, when you didn't know what they were, you had to clone them? So, you purified a lot of RNA from your cells and then you used to enzymes to ligate small adapters on each end of the molecule. And you could clone it into a vector and sequence it. And we did that in the early days. So, you sequenced and cloned a lot of small RNAs, then you tried to find out is this a micro-RNA. Or is it just some debris of other RNA molecule in the cell? So, once you had all these sequenced and cloned and you tried to figure out what they actually were doing. And now once now that we know what micro-RNAs are actually in a cell, you can just use array types or PCR to follow their expression.
Jordan Ruggieri 03:56
What challenges or obstacles have you encountered studying micro-RNA ?
Guy Novotny, PhD 04:02
To figure out what they are doing it's very difficult. I mean, it's easy enough to clone and sequence them, you might say, and also to investigate their expression. Once you know they're there, the arrays and the qPCR assays are pretty well functioning. But once you try to figure out what they actually regulate, they bind to messenger RNAs complementary. But since the sequence they use to bind is only about 6,7,8 nucleotides long, the potential targets they can bind to are hundreds, maybe thousands. Maybe so when you have a micro-RNA that you're interested in to find out what it's actually regulating is very difficult. Actually, sometimes you go the opposite way around, you have a protein you know is regulated in some way, but you don't know how and then you try to figure out are there any micro-RNAs that can bind to its messenger. And then you check, does it actually do that? So, you maybe destroy the binding site on the messenger RNA and see if this helps its expression and stuff. So, it's, but this is a, that's a major thing. It's easy enough to follow the expression of the micro-RNAs, but to figure out what they're actually doing is the major challenge.
Jordan Ruggieri 05:09
How do you actually, you know, look and see if a micro-RNA is going to have an effect on translation? Is it a, is it a computational? You're looking through databases and identifying possible sequences? Or is it, is it more experimental?
Guy Novotny, PhD 05:27
You would initially do a bioinformatic approach. Use computation to figure out which are your likely targets. And then if you know what cell you're looking at, then you can see are these likely targets, anything that's expressed in the cell. Is it something you're interested in? So, you can do things like that. A lot of work and a lot of negative data, you might say. Too big a chance to, to fail when you have so many options for what, for what you're looking at. It's a difficult field. I switched from, from pure research to clinical lab work about 10 years ago. So, I've also continued some of my micro-RNA research with students as a supervisor. But basically, I've been more focused on the clinical aspects of molecular biology for the last maybe 10 years or so.
Jordan Ruggieri 06:17
What exactly are you looking at? Is it is it along the lines of micro-RNA still? Or is it is it completely different field on the clinical side?
Guy Novotny, PhD 06:25
It’s different now because the diseases I'm looking at the micro-RNAs haven't really shown to be that important in the decision making of how to go forward. In some in some diseases micro-RNAs are important and are also likely, might say, prognostic or diagnostic factors in the disease, but what I'm looking at, it doesn't seem to be that important. When I do research now on micro-RNAs it's basically for fun and for developing the field.
Jordan Ruggieri 06:59
Awesome, nothing like doing research for fun.
Guy Novotny, PhD 07:02
In your spare time, on your own.
Jordan Ruggieri 07:04
Switching a little bit here. How, you know, we love digital PCR here at Absolute Gene-ius. How do you utilize digital PCR in your research?
Guy Novotny, PhD 07:13
So, I've been following the digital PCR research for a while, but I haven't until recently had access to a digital PCR machine. But now that I have tried it out is you might say you can do the same with qPCR that you can with a digital PCR machine, but the precision of your data that you know, the number of copies you measure with a digital PCR is a lot more certain the number than if you use your qPCR where the efficiency of your reaction is so important and difficult to control. It's so much more easy to use a digital PCR and then get a result you can trust.
Jordan Ruggieri 07:54
Yeah. Uses the same chemistry and makes the makes the assays kind of easy to use between the two. Are there any examples or any situations where you might use digital PCR over real time PCR? Is it just that precision element? Or is there any effect that you know looking at micro-RNAs, digital PCR is a better technology for you?
Guy Novotny, PhD 08:22
Yeah, the digital PCR is very convenient when you want to multiplex, is the word. So, if you want to test for several genes or several targets in the same in the same well then then the digital PCR is it doesn't really care what the efficiency of your reaction is. So, it'll tell you exactly how many copies you have even if you have two or three or four different targets you're looking for with different colors. If you do a qPCR there the efficiency of each of the reactions, the it will be hard to keep them 100% efficient and then the numbers you get will be skewed compared. So, when you want to compare them you know that it will be a wrong comparison. Otherwise, you have to run each of the targets in each well, in different wells, and then you have the problem that you have technical variation that's difficult to control. So, it makes it a lot easier using the digital PCR just put everything in the same well then you know you have the same amount of material there and everything no matter the efficiency it gives you the correct data.
Jordan Ruggieri 09:26
Do you often multiplex with micro-RNA, or you're often looking at multiple targets?
Guy Novotny, PhD 09:31
Initially we, you always use multiple targets because you need to control for the technical variation of the RNA quality of your samples. You always have to when you're, if you're looking at a lot of different samples and you then you need to know you are putting 50 nanograms of RNA down there to make your cDNA but is it 50 nanograms of good quality cDNA and RNA. So, you need to, you need a control gene always to compare your micro-RNA of interest to. If you don't do this comparison correctly, if you don't have this gene to compare to, then you basically can't use your data for much.
Jordan Ruggieri 10:09
Yeah, that makes a lot of sense. Do you ever find situations where you might use both real time PCR qPCR and digital PCR, together? Or kind of go back and forth between the two technologies?
Guy Novotny, PhD 10:23
You can go back and forth. The digital PCR has a limitation of overloading, you might say. If you have a lot of material, the whole, if you don't know anything about digital PCR, it does use a sort of statistics to assume how many copies you have spread out in your plate or in your droplets or whatever, you're dividing your sample into all the small partitions. If there, if there are no empty partitions, then you can't do any calculation based on this Poisson distribution. You don't know how many copies you have to use, you have to have a certain amount of empty wells before you can actually make your calculation. And so if you have a lot of material than your digital PCR, it will be overloaded. And it'll be difficult to do your calculation. So, the qPCR can have a wider range. So, you can have a lot more material in the qPCR.
Jordan Ruggieri 11:20
We do see that often where, depending on the dynamic range you need and your exact parameters of your experiment and the answers you're trying to get, utilizing both technologies can be helpful sometimes.
Guy Novotny, PhD 11:33
For sure.
Jordan Ruggieri 11:34
We get comments all the time, about the cost effectiveness of moving over to digital PCR. And I'm curious if you've run into situations where in qPCR, when you set up real time PCR, when you're running things in triplicates, you have to use standard curves, right, do you see an increased number of reactions that you have to run versus say a multiplex experiment on digital PCR?
Guy Novotny, PhD 12:02
Yeah, so we do, we do a lot of qPCR as standard and we've been looking into to switch, switch it over to the digital PCR to see if it's faster, better, easier. The more high-throughput assays that are where the standard curves business is a smaller part of the whole assay because you do it only a little bit but you do a lot of runs, it might be a little a little bit less cost efficient. But still in the very sensitive analysis we do, we have sometimes actually 12 replicates. We have six genes with our gene of interest and six with the different reference genes. And I have recently been doing some tests and I can reduce these 12 replicates down to one with digital PCR. So we just put it all in one in one reaction. So, that's a lot of time saved. And it seems to work just as well, the same sensitivity and the same precision.
Jordan Ruggieri 13:02
That's awesome to hear. Yeah, really, really cool. We talked a lot about some of the details of micro-RNAs and what they what they do and some of your workflow. What is the ultimate outcome that you're hoping to achieve with your micro-RNA research?
Guy Novotny, PhD 13:15
With the micro-RNA research, I was, we're basically hoping to better understand the disease you're looking at and then of course, follow up with that to maybe visualize what sort of treatment or cure you could do. Often this is a very long term work path. A lot of times you're just hoping that you can discover something that can maybe help place a patient in a specific group so you know what treatment they should have. Or maybe just get a little bit of better knowledge of how a protein is regulated to just, for like adding to the big get pile of data that that's existing out there, and then someone else will, will build on it, and then in 20 years maybe someone who views your research to make something useful. But that's, that, you might say is basic research. We also do, you might say, practical research where you don't ask a super large question, but just say is can we do something a little better and a little faster and is there something specifically right here that if we change or do a different analysis can get a better answer. That's also quite interesting. It's to do because sometimes you can produce something that other people can use almost immediately. But of course you can always hope for the Nobel prize. But often we hear that the people that get a Nobel Prize idea, they do that when they're around 20 or 30. And I'm too old for that now, so it's my chance is gone. I don't have such hopes anymore. I'll just add a little bit of data to what's out there and then hope so hope someone can build up on it.
Cassie McCreary 14:59
Taking a quick break from our conversation to tell you about Applied Biosystems™ QuantStudio™ Absolute Q™ dPCR system. This instrument enables quantification of your targets without the need for standard curves in only 90 minutes. Digital PCR can be as simple as preparing your samples, loading onto the plate, and running the instrument.
Jordan Ruggieri 15:18
Unlike other digital PCR systems, the Absolute Q dPCR instrument does not use emulsion or other droplet-based methods to compartmentalize reactions. In fact, the microfluidic array plate (MAP) technology enables consistent delivery of more than 20,000 micro-chambers. It's a great solution for anyone looking to quantify gene targets.
Cassie McCreary 15:40
And Thermo Fisher Scientific has a suite of dPCR assays for applications like AAV viral titer quantification, liquid biopsy analysis, and wastewater surveillance. You can learn more at www.thermofisher.com/absoluteq or visit the Absolute Gene-ius webpage. Again, that's www.thermofisher.com/absoluteq or visit the Absolute Gene-ius webpage.
Jordan Ruggieri 16:07
The Applied Biosystems™ QuantStudio™ Absolute Q™ dPCR system is for Research Use Only. Not for use in diagnostic procedures. Let's get back to our conversation.
Cassie McCreary 16:20
Time for Cassie's Career Corner, Guy. Welcome. This is the part where we talk about your, just, your career and fun stories and all kinds of just goofy things. I was hoping you could kick us off. I know you've talked about this just a little bit. But I'd like to go a little bit deeper, just kind of an overview of your academic studies and kind of the various steps throughout your career.
Guy Novotny, PhD 16:44
I started out working with bacteria and ribosomes, ribosomal RNA, because if you don't know but a lot of the if you're sick with the with the bacteria without an infection, you will eat some penicillin and some pills and it'll kill off the bacteria then you get well again. But a large amount of what you eat of medicine, it actually targets the ribosomes, the ribosomal RNA, and the ribosomes if you don't know what that is, that's actually the machinery inside the cell that makes all the proteins. And a lot of the resistance mechanisms so if you have a bacteria that's resistant to the medicine you're eating, it's actually often caused by either a point mutation in the ribosomal RNA, so the antibiotic can't bind to it and prevent its function anymore. Or it could be some cellular pump that pumps the medicine out again, but a lot of times it's actually just the ribosomal RNA that's mutated, and then the antibiotic just doesn't work anymore. So, antibiotic resistance is a very important field to do research in so that was that was interesting to do in the early days. But again, after you've done it for maybe four, five, six years, growing E.coli bacteria, doing this research, sometimes you want a change of scenery. So, then I got the opportunity to work with the human samples, human cells. Well when I was finished actually with my masters then I worked with, did a PhD project at a hospital working with the testes cancer and leukemia, a split project with the collecting it together with say the micro-RNA. So, I was looking at micro-RNAs in testicular cancer and with the with the leukemias, B-cell leukemia. Not much was known at that time. So, we started out just basically cloning stuff looking what was there trying to describe it. That took a couple of years. Then actually I started working with diabetes. Because in Denmark, where I'm working the one of the big companies is Novo Nordic and they have a lot of research on diabetes. So, there's a lot of money funneled into diabetes research. There was a group that wanted to look at micro-RNAs and diabetes. So, I did about three years of a postdoc position describing and looking at the micro-RNA function in diabetes. That was pretty interesting, but then funding is always hard to get so and when you do a career change as often as I have from different areas, you sort of might say you lose the momentum you had in, in you leave behind piles and mountains of work that don't get finished, that doesn't get finished, and then you start up again from scratch. Takes some time to get going. So, after changing from the ribosomes to the testicular cancer and leukemia and then over to diabetes my CV, you might, I say my publication list wasn't as large as the people I'm competing with that's been staying in one area. So, I had to look for alternatives to continue my work. And that was a job at a hospital as a molecular biologist to validate assays to interpret data to develop new methods. I've been going from more and more basic research and basic data to actually working with patients that sometimes you can find stuff that really, really helps them. So, you're going from, you know, basic to practical research. You're doing, you feel like you're doing more day to day decisions that are affecting people immediately.
Cassie McCreary 20:22
Your career path is it's a pretty unique one. A lot of the guests that we've had on the podcast so far tend to, they've gone into an area, and they've stayed and focused on that area for quite a length of time. So, seeing how you've jumped from a few different areas of research and then to clinical. It's interesting to see that path and I think that's something that's really unique for probably our listeners to hear about. So, thank you for sharing about that. How is it that you decided that you wanted to go down the route of science and molecular biology? I mean, did you always just love science? Or is it something that you kind of discovered, as you made your way through? I mean, what's that like for you? What does that path look like?
Guy Novotny, PhD 21:03
I've always known that I wanted to go to the university and study some, basically, genetics, I think was what I wanted to do. I have a big passion for science and science fiction and fantasy literature. And when I when I'm reading this, then I'm thinking, "Ah, yeah, I want to I want to clone and do mutations." And it was part of my interest.
Cassie McCreary 21:27
I like that. That's interesting. Yeah, that's good. I mean, it's, it's good to know from the start. Some people kind of go through their career, and they feel maybe a little bit lost. And they feel like they have to try a bunch of different things before they find the thing that really makes them happy. But it sounds like for you pretty early on, you're like science is it. And that's great.
Guy Novotny, PhD 21:43
Some of my decisions were based on circumstance. So my work with the bacteria and ribosomes, it was just because there was an open position in that lab to do my bachelor's and master's degree. And my work with the micro-RNAs, it was basically because the one I was, the person I was working for knew, had a friend that wanted that. So, that's where I ended up. And my diabetes change was because my wife was working with a colleague that knew that they needed someone to work with micro-RNA, so I switched over there. So, it's been, I have switched from area to area. But the science was my focus in the RNA, but the area of the science was basically by chance.
Cassie McCreary 22:24
I think that's important too, though, because another sort of theme that we've pulled out over time on the show is like there's an, there's an importance that you, of being open to new possibilities that maybe you hadn't necessarily considered when you're sitting down, and you're kind of picturing what your career path looks like. So, I think that's kind of another really good testament to that. You can plan all you want, but you never really know what direction things are going to go. Would you say that there's been a great degree of importance in having a network and mentors throughout your career?
Guy Novotny, PhD 22:55
It's always helpful and it's always useful to have a large network to ask and get guidance from. You never know enough yourself. There's always a method that you've never heard of, or a connection between data that you won't see if you don't have enough people to ask. Maybe with these AI days coming up, you can just ask Chat GPT how this all is connected. But in the olden days, you needed to ask around. You didn't have enough information to get it correct.
Cassie McCreary 23:27
That's a good point. And if I'm remembering this correctly, you're from the U.S. originally, right? I want to say Minnesota for some reason. Do I have that correct?
Guy Novotny, PhD 23:36
That's correct. Minnesota.
Cassie McCreary 23:37
At what point in your life did you move? Was it around when you were pursuing your studies? Or was it very early in life?
Guy Novotny, PhD 23:42
It was when I was around 10 years old.
Cassie McCreary 23:44
Oh, okay.
Guy Novotny, PhD 23:44
So, my mother's Danish and my father's American.
Cassie McCreary 23:48
Okay.
Guy Novotny, PhD 23:48
When I was around 10 years old, my mom wanted to move back to Denmark.
Cassie McCreary 23:52
Okay. All right.
Guy Novotny, PhD 23:54
We packed up and moved.
Cassie McCreary 23:55
Bye Minnesota.
Guy Novotny, PhD 23:56
Yeah.
Cassie McCreary 23:56
Interesting. Okay, very cool. A lot of our listeners, we're hoping, are kind of earlier on in their career or they may be finishing up school or they're partway through their studies. So, what would be a piece of either career or even just educational advice that you would have for them?
Guy Novotny, PhD 24:11
If you want to, I don't know if it's to succeed, but if you want to have fun, then you have to do something you're interested in. Because you don't, I don't think you can really avoid using a lot of hours. Unpaid, unpaid, hard, late hours when you're doing research. And if you don't like it, then it's not going to work. So, your work has to be your hobby. If you're too stressed out and doing stuff you don't want and using so many hours on it, then it's not going to work.
Cassie McCreary 24:37
Okay, yeah that's great advice. Thank you. And so, we like to ask this question of everybody who's on the show. You don't have to answer it if you're not comfortable, but we love hearing these stories. What is your biggest mistake, or mess up that you have made in the lab?
Guy Novotny, PhD 24:54
Well, I think most of us have taken out the enzyme box, the little freezer with all enzymes in to keep them cool. And you use the enzyme. And then you put it, so it doesn't get, you know, you don't want to heat it up, and you put it back. And then of course, there's always one that forgets to put the whole enzyme box back in the freezer. And the next day, you didn't just ruin the one little tube you need, but the whole stock of everything. And it's and then the professor comes and says "That's coming out of your wages," but it can't be paid for years. So it's, it happens once in a while, then you have to restock the freezer. But it's you're told that you're not supposed to do that again. You just you have to bring cake the next day.
Cassie McCreary 25:36
You have to bring cake. Alright, well, then that's an even bigger piece of advice, everybody, if you leave out the enzymes, make sure you bring cake the next day. I love that.
Guy Novotny, PhD 25:42
I think, oh yeah, what have I done? Well, I had a, I think I had two or three months during my PhD, where I just always got a PCR product up in my water sample. My non-template control. So there's, there was a contamination somewhere. I just don't understand where it's coming from. You're just rerunning everything, changing out my stocks of primers, water, all the buffers, everything. And then can I know what's myself that's contaminated it because it's I'm the only one working with this PCR product, this gene. So, it's only myself, I could have done something with it, I couldn't figure out what it was. And finally, after a couple of months, it disappeared. And that was when the enzyme was used up. So, somehow my PCR product had come into the enzyme stock. So, every time I was running the PCR, I'm always getting the kind of contamination. That really, and I didn't understand where, I mean, you're just thinking, how did it how do I put a PCR product down in my in the enzyme stock. But after that happened, I'm very careful with it with the enzymes because it's I mean, it was months of work, right? You wouldn't trust any data, you just had to throw everything out until it went away. And it didn't went away until enzyme was used up.
Cassie McCreary 26:53
That's awful.
Guy Novotny, PhD 26:55
Be careful with the with the enzyme. I think that was before there was something called a filter pipette tip. So, when you were pipetting any of these plasmids, you have them there are millions and millions of copies slowly went up into the pipette. And then when you're pipetting, next time, if some of it fell down into the enzyme, I believe that must. After we got the filter pipettes it wasn't so much of a problem.
Cassie McCreary 27:16
Wow, that's a challenge. I think, I think one of the things that is like, so overlooked about careers in research and all that is the massive amount of patience that you have to have. Because I would have never been able to get through, I would have like, “I don't know, it's a mystery. And now it's two or three months, and I can't do this.” How did you decide to keep working despite that?
Guy Novotny, PhD 27:44
If have an experienced professor or day to day supervisor, you always have more than one project you're working on. Because there's always, it's always something's going to not work and you're going to go hit a roadblock and you're not going to solve it for weeks and months, maybe. So, you always have two or three different projects you're working on. So, at that point, I think it was it my testes cancer project that sort of got put on hold, I couldn't get it. But then I had the leukemia project. So you did, you do more than one thing, if you're just sitting totally blocked for months on one thing, it's, it's going to, it's not going to be good. But if you tried to have several different projects, working some that you might call the simple bread and butter projects that are certain to work, no matter how much you fail, and then you have more and more risky projects that if it works, then you're going to be really happy. And if it doesn't work, then no problem.
Cassie McCreary 28:36
Interesting. That's good to know. Alright, so now that we've covered some of the more oops side of things, what would be your most exciting or proudest or happiest moment in the lab?
Guy Novotny, PhD 28:47
When you sort of discover something maybe new or unknown. Or you have like something that everyone knows is correct and then you find out, it's actually not correct at all. I mean, then you think, “Okay, I can, I've learned something new, and I can try to teach it to someone.” And there have been a couple of moments where also when you're like developing new methods, and you get them to work and then you can say, "Hey, if people use this method is you're going to save time, save money, maybe get a better data." There are a lot of different small things I think have made me pretty, pretty happy and made me proud and laboratory that I got something changed for the for the better. But some of the stuff I've been working on this, for example, like RNA I've been working a lot with RNA and people are pretty, you got to be careful with it because it can there's a lot of enzymes that break down RNA, they're all over the place. So, if you just leave your RNA around it will be broken down and degraded and useless. But actually RNA in itself is very, very stable, but people don't really realize it. So, if you have pure RNA and put it in water and just put it in the refrigerator, or then leave it on the table, it's not going to break down instantly. It can actually last quite a long time. And what at one point, I had put my RNA in a PCR machine, heated it up for 40 cycles up to 95 degrees and down again. And then I run it on a gel, and it was still perfectly intact. So you could so it's that surprised me that you could easily basically boil your RNA for models will 30 minutes in your PCR buffer and nothing happened to it. Still intact.
Cassie McCreary 30:24
Oh, yeah, that's a lot more resilient than expected. Yeah.
Guy Novotny, PhD 30:27
Yeah. So, when sometimes when you see these things, you'll think, "Hmm, is everything I know correct? Or is it wrong?" But there's basically I think what I've found out was if you don't have RNAases, the enzymes that break it down in your solution, then it's stable, much more stable than you would expect.
Cassie McCreary 30:46
You hit on an interesting point with that, too, because you said, you know, is everything I know, correct or you know, whatever. But I think that isn't that kind of the root of science anyway, you think you have things figured out, and then a new discovery comes along, and it totally shakes things up. So you make a good point with that. Now, I know you spoke about how in research, you'd have to find a labor of love. There's a lot of you know, unpaid hours, there's a lot of time spent in it and that type of thing. Do you have interests outside of like, what do you do outside of work to give yourself a mental break from some of this?
Guy Novotny, PhD 31:15
I do a lot of reading when I can. Still the fantasy and science fiction genre and, and a lot of computer games.
Cassie McCreary 31:23
Okay.
Guy Novotny, PhD 31:24
But I do have, I do have five children.
Cassie McCreary 31:28
Wow. Okay.
Guy Novotny, PhD 31:29
So, my wife is also a researcher. So, we all have long hours and a lot of, we don't have much time.
Cassie McCreary 31:36
I would say not.
Guy Novotny, PhD 31:38
When I play computer games, they have to be something that you can start and then stop at any time and then continue again. So, like you might say online games against other people, that's not going to work. But turn based games where you can play against a computer and just stop whenever you don't have time. That's going to work. And books that you can just close and start again. That's fine too.
Jordan Ruggieri 32:02
What are your favorite books?
Guy Novotny, PhD 32:04
I do have some science fiction books from a writer called Peter Hamilton, I think his name is. And what I've been reading now I've been reading. I guess the last books I've been reading was maybe an American writer called Sarah Maas. The series called Throne of Glass, or something I just found those.
Cassie McCreary 32:31
Oh, I've heard of those.
Jordan Ruggieri 32:33
Okay.
Guy Novotny, PhD 32:33
And they're very nice. I've been reading those.
Jordan Ruggieri 32:35
Awesome. I'm a big reader as well. Big fantasy and sci fi reader as well. So, when you get the chance, you have to check out Brandon Sanderson. You'll like his stuff too, if you haven't read anything from him. So.
Guy Novotny, PhD 32:47
I haven't heard. I haven't heard from him, no. But I'll check it out.
Cassie McCreary 32:53
That was Dr. Guy Novotny, molecular biologist at Herlev Hospital in Copenhagen, Denmark. Thank you so much for joining us for today's episode of Absolute Gene-ious. It was produced by Sarah Briganti, Matt Ferris, and Matthew Stock. With more great science around the corner in future episodes, stay curious and we'll see you next time.
Cassie McCreary 33:12
That's it. The whole next interview is just going to be done in song and preferably interpretive dance even though people can't see it. We should get matching unitards. With little DNA's on it.