Hello everyone. Thank you for spending some time with Promega today. I'd like to welcome you to our webinar, Simplifying Lab Work: A Guide to Laboratory Automation Using liquid Handlers. My name is Eric Vincent, I'm a Senior Product Manager here at Promega. I'm specializing in high throughput extraction, and I'd like to begin today by telling you a little bit about the platform and how to interact with it. So... There we go. So, oops. Before we begin, I'd like to cover a few house keeping items. On your screen there are multiple windows, and all of these windows are movable and resizable. So feel free to reorganize the desktop space so that you can better see the webinar and interact with it. We have many ways to be interactive. You can submit a question at any time during the webinar, and we'll answer these during the live Q&A question, Q&A session at the end. We'll also be chatting with you if there's a simple question we can answer just in chat. The resource library that you can see is also filled with some useful materials, including a copy of today's presentation. So feel free to download any resources or bookmark any links that you may find useful there. Also check out our Lab Resource Center, which is on promega.com which has a lot of information on automated nucleic acid purification tasks, etcetera. You can also view an on-demand forum on nucleic acid extraction. We had several experts on last month from our automation team. And we'll also have a short survey after the webinar. So please take a moment to answer those questions because they really help us as we schedule, design, and present additional content, because we really want it to match your needs. And then finally if you're liking something you see, please feel free to give a reaction to the presenter to know if you enjoyed a slide or the presentation itself. You're welcome to share this webinar with others if you think they would be interested as well. But with that, I will move forward and introduce our speaker for today. Today's presenter is one of our applications scientist, Dr. Sarah Teter and she works in our applications team where she helps determine new uses for our products and for our customers, and also works with our automation team to help automate the nucleic acid extraction and other tasks on laboratory automation systems. With that, I will turn it over. And Sarah, thank you for presenting today. First thing, sorry for, thank you for the introduction. Thank you, Eric. And so yeah, like Eric said, I am an application scientist and I really have a passion for high throughput automation and making it work with Promega's chemistries. And so without further ado, I will jump right into the webinar. So if you guys are familiar with Promega, you probably recognize us as primarily a reagent company. So we provide reagent tool kits across many different kind of areas of science, including cell biology, DNA, RNA and protein analysis, drug development, human identification and molecular diagnostics. But what we know is that by pairing these powerful reagents with automation, we can really empower scientists to do more awesome science. And so we have a whole team dedicated to automation here at Promega, and that team is our field support scientists. You'll get to meet one of those scientists later this morning. But I want to mention before I get into any of the meat of the talk, that we do offer automation support for Promega products. The support is free of charge and it's also platform agnostic, meaning that we aren't going to try to push you towards a particular instrument. We really want to help you find the instrument that is going to work best for you. And so with that, let's jump into the actual content of this webinar and talk about what my goals are to bring across through this talk. First, I want to demonstrate, of course, how automation can make your life easier. And we'll then also use a couple of real-life automation examples to map out a manual process to be able to talk about how to translate that to an automated process, and also describe some basics about automation equipment that's available on popular liquid handlers. So in terms of laboratory automation, there are really 3 levels of laboratory automation. Of course, the one we're probably all familiar with, at some point in our careers anyway, is no automation. The second level is really, it's called modular automation. And then working up from there is integrated automation where we kind of join together different modular pieces into a whole automated workflow. So let's talk a little bit more about each of these. Of course, with no automation, this is just kind of the status quo and there is kind of this perception that no automation is inexpensive. But if we think about having to spend your time pipetting in the lab, what is that preventing you from doing kind of more ambitiously in terms of your science? So I don't know that it's really inexpensive, but it is just kind of what is set in stone historically. In terms of the cons, no automation is definitely time consuming and tedious. At some point, when you have enough samples or assays that you need to process, you can't, you can't do more yourself, and at some point you get limiting in terms of the number of actual scientists you can have in your lab, so it's not scalable either. Finally, just due to the human element with no automation, of course there's a tendency for errors, particularly if you're doing repetitive processes over and over. Stepping up from no automation is modular automation. And with modular automation, what we're looking to do is just to automate the most tedious aspects of a workflow. So it's not completely hands off. There's still going to be some user intervention with modular automation, but the goal is really to decrease hands on time for those time consuming tasks. We're also looking to reduce errors and increase reproducibility and accuracy. And finally, it's a way that you can reduce risk in the lab. And when I'm talking about risk here, I mean a couple of different risk factors, including things like repetitive motion injuries from pipetting, and also potential exposure when you're handling infectious materials in your lab, if you are processing those types of samples in your lab. Some of the cons though with modular automation are that it still requires some user intervention, as I mentioned. There are initial and ongoing expenses and these expenses can range from moderately to moderately-high expense depending on the exact system that you need. And then you need to factor in about 10 to 15% of the initial cost of the instrument for annual service contracts. So there are those ongoing expenses to keep those instruments maintained. You also need to develop expertise. So if you don't have experience with automation and liquid handlers in your lab, you will need to target somebody or a couple of people to be able to develop that expertise to be able to support this instrumentation. And finally, I just wanted to mention that you will need to factor in redundancy in your processes for being able to have downtime on the instrument for maintenance and any potential instrument failures. The last level of automation here is integrated automation. And with integrated automation, like I said, that's kind of a step beyond modular automation. And the goal with integrated automation is really to automate entire workflows. By automating the entire workflows and minimizing hands-on time, you can definitely greatly reduce errors, increase reproducibility and accuracy again, and further reduce risks to your lab staff. The cons, though, with integrated automation, are those initial and ongoing expenses. Considering that each of these kind of modules within the integrated system can cost tens to hundreds of thousands of dollars, it can be very expensive to get set up with the system and again those ongoing expenses due to service contracts. Here, kind of the level of expertise that you need to develop for an integrated automation system is even many steps above kind of what you need for modular automation, because you need to have expertise not only on one system, but across multiple systems and you need to have the ability to link those systems together so that they can communicate and function together. Again, just because you're linking several pieces of equipment together, there's the need to factor in time for maintenance and potential instrument failures. So, obviously today as an automation webinar, I'm not interested in telling you about no automation. And really, integrated automation is a step beyond what I want to talk about today. So the focus that we're going to have today is specifically on modular automation. So to jump into the modular automation, we want to talk about first some of the capabilities of the lab automation systems because this is going to help you understand what types of processes are best amenable to automation. So when we're talking about liquid handlers, firstly, one of the big things that we're looking to automate is being able to pipette liquids. We also can use barcode readers for sample and reagent tracking, shakers to mix samples, centrifugation is possible on certain systems, temperature control, being able to manipulate plates around the deck of the instrument or tubes, and then there are some tube capping and uncapping devices. And so factoring in all of these different capabilities of lab automation systems, there are many workflows that can benefit from automation on a liquid handler. And this is definitely a non-exhaustive list. I'm sure if you guys put your minds through it, you can come up with a lot more potential workflows. But these range in kind of complexity from really easy processes like serial dilution, tp things like NGS library prep and forensic sample analysis, where you would have sample extraction paired with quantification and normalization, and then finally setting up those STR reactions. But I want to take two of these workflows and use them as examples that we will talk through to explain to you how we at Promega kind of approach automation when we are talking with a customer. The first example that I'll be leading you through today is one in the realm of cell culture and analysis. and then the second example will be in nucleic acid extraction. So starting off with this first example of cell culture and analysis, and again, these were real customers that we worked with. Of course, I'm not going to identify exactly who they were here. I'll tell you a little bit about who they were and what they were doing. This first customer was part of an academic molecular screening lab and they were interested in testing novel compounds for their effect on cell viability. So they already had a back catalog of about 10,000 compounds, and they were adding additional compounds through collaborators on a weekly basis. Each of those compounds would be screened over a range of concentrations and against multiple cancer cell lines. And they were looking to kind of miniaturize to a 384 well format because that offers savings and time in terms of reagent cost. There are many reasons why this this scenario is amenable to automation. Firstly is just the large number of compounds that they're working with. But then that large number of compounds is going to be expanded across multiple concentrations and cell lines. So you have many, many assay wells for each compound that you're testing. And then working in this higher density plate format, if you guys have worked at manually pipetting into a 384 well plate, you know that that's really easy to lose your plate when you're pipetting into those plates, particularly if you're adding multiple reagents to each well of that plate. So really, it's best to use the 384 well format with automation. And so as I mentioned, they were interested in looking at cell viability. So the Promega tool that was kind of the one that they selected for this work was the RealTime-Glo MT Cell Viability Assay. This assay is a luminescence-based assay, where the luminescent signal is proportional to the number of viable cells in each well. And the kind of big benefit of this assay is that it is a kinetic assay, meaning that you aren't just taking a single endpoint read, you're able to read cell viability at multiple time points over a time course for up to 72 hours. And this is really powerful in this scenario where we are looking at novel compounds when we're not sure exactly when we might see an effect on viability or if we'll see one at all. And so, using this type of assay where we can take multiple time points allows you to gather more data per well of the assay, more data per well in the assay plate, as well as just allow you to get some really robust data. So pairing that RealTime-Glo Cell Viability Assay with the an instrument from Beckman Coulter, called the Biomech I series. This actually had already been selected by the customer before we were called in to help support them. And again, I just want to emphasize that the support of our field support scientists is instrument agnostic. So we will work with any instrument that you have or help consult with you to figure out which instrument will be the best for you in automating your Promega workflows. So when we have that kind of general workflow, the first thing that we need to do is break that down. And so we can break this cell culture analysis protocol into 3 broad steps. The first step is going to be to prepare and plate the cells. Then we'll prepare serial dilutions of the compound and add that to the cells. And finally we'll prepare the RealTime-Glo reagent and add that to the wells so that we can monitor cell viability. This level though of the breaking down the workflow is not sufficient to understand how to automate the process. So we're going to talk a little bit more in detail about the specific things that we need to consider, including things like where the reagents and samples are being stored? What types of labware are being used? Are there specific plates, reagent reservoirs and tubes that you're using in your process? What types of consumables you're using? So these are things like tips, what size tips do you need for your process? And how many will you use for each plate that you're processing? You also need to consider the scale of the volumes to be pipetted. Are we pipetting nanoliters or milliliters? Or in this case we're pipetting microliters worth of volume? And finally, once you've mapped out your process, you really want to think hard about what is most important for you to automate, versus what is not so difficult or time consuming to just prepare manually. And so kind of breaking each of these broad steps down into more detailed steps for preparing and plating the cells. We'll start with our cells in a T-75 culture flask. And then the first step after that, if your cells are an adherent cell line, are to trypsinize those cells and split some off for passage. Then you'll take the remaining cells, count them and then dilute them to the appropriate number of cells per mil, and transfer the cells to the appropriate wells of a microtiter plate. So now we have our cells plated. The next step is going to be to prepare the serial dilutions of drug and add that to the cells. We'll remove our compounds from storage, in this case, the customer restoring their compounds at -20. So obviously they might need to allow those to thaw before they prepare them. They need to somehow document the compound IDs and then they will start the process of making serial dilutions. So typically the first step of serial dilutions is to aliquot your diluent into the wells or tubes that you're making, preparing the serial dilutions in. And then, for example, preparing that first serial dilution, you would aspirate from that stock tube of compound, dispense it into the first dilution tube, pipette to mix, and then we'll be pipetting from first dilution tube to 2nd, 2nd to 3rd, and so on until you've prepared as many dilutions as you want to assay. So that was making a serial dilution for a single compound. This customer was assaying 4 compounds per plate. So we need to repeat that serial dilution process for additional compounds. After we have the serial dilutions for all the compounds, we'll transfer the compound serial dilutions to the microtiter 384 well plate. The last step of this, once we have our cells and drug plated together, is to add the reagent so that we can monitor cell viability. And so the first thing we're going to do is remove the reagent from the freezer and equilibrate it to room temperature. Then we need to prepare dilution of the reagent. And finally add that diluted reagent to the wells of the microtiter plate. So now if we kind of consider the whole workflow that I've laid out and think about what capabilities we need to incorporate into the system, or what can we incorporate into the system, to automate this workflow. Of course, we definitely need pipetting, the ability to pipette liquids. There's no way to get around that with this process. We can also use, could potentially use, barcode readers to track those samples or those compounds as they're being plated for each of those assay tests. We could potentially use shaking to do some of our mixing steps. Depending on how we set up the assay, we might need to move plates and tubes around the instrument. And we could potentially use tube capping and uncapping devices. There are two pieces of equipment here then that I've left out. Obviously we don't need centrifugation, and temperature control is something else that we just don't need for this particular workflow. Now looking at the types of equipment that we actually want to include, we decided in consultation with the customer that sample and reagent tracking wasn't necessary. They were going to track those compounds manually to keep track of where what assay plate had those compounds in it. We also eliminated using shaking for the mixing. It really just is not the best way to mix in this application. We get much better mixing, particularly of the serial dilutions, by employing tip mixing. We had no need to really move plates or tubes around the deck of the instrument. So that was another piece of equipment that we eliminated. And finally we eliminated the use of the tube capping and uncapping equipment. And the reason why is with those tube capping systems, you need to use specific tubes that are compatible with those systems. And so if they opted to use the tube capping system, they would need to require that their collaborators send the compounds in specific tubes. And that would restrict how they receive samples. And really we're just functionality that we are going to incorporate into the workflow and that is pipetting liquids. With liquid pipetting there are a couple of ways that we can achieve this on liquid handlers. We can either use pipetting channels or we can use a pipetting head. With pipetting channels, these are typically provided on an instrument in sets of eight, and they have the ability to move independently in the Y&Z axes. So the Y-axis is from back to front of the instrument, and Z-axis is up and down in space. They are very flexible in terms of their utility on these systems. So they're useful in cases where we might need to pipette different solutions across the plate or where we need to do hit picking, which would be something like sampling individual wells on a plate. Or if we were sampling from non-automation-friendly tubes where they aren't arrayed in that kind of 8 by 12 format that we're accustomed to in automation. Pipetting channels are also useful in cases where you have, or you want to use, minimal reagent dead volumes. And when I say reagent dead volumes here, this really speaks to the minimal residual volume that's required to avoid aspirating air. And this goes to waste. Or this reagent goes to waste. And so with the pipetting channels, then, where we have kind of these limiting reagents, we can make the best use of those reagents by using the channels. With the pipetting head, then, these can be provided in 96 and 384 well formats. And really the usefulness of these pipetting heads is that they are very fast. So in cases where you need to stamp reagent or sample across an entire plate, these work great. Because where with the pipetting channels you would need to pipette 12 times across a 96 well plate, with a pipetting head you could simply do one pipetting step and have that reagent pipetted to all those wells. It's also useful in cases where you need to do replicate plating. And replicate plating would be something like where you start with DNA samples in a 96 well plate and then perhaps you're setting up qPCR reactions and in a 96 well assay plate. You can simply transfer from that DNA sample plate all 96 samples into that assay plate in one quick step with the pipetting head. The kind of drawback to the pipetting head is the higher reagent dead volumes. So if you think about pipetting with this reagent head, we have to use a reservoir that's much larger to be able to accommodate all of those tips going into the solution. So they have much higher reagent dead volume. So this isn't the solution that you'd want to use if you have small volumes of reagent. So I want to take a pause here and talk about basically choosing a good instrument supplier. And I want to emphasize that you don't want to get pressured into buying the Cadillac of automated systems where you have every possible piece of equipment on that system. It really is important to know what methods and tools are best for your process and workflow. You want to avoid adding additional equipment to the platform that's not going to be crucial for your workflow. So in the example that we've just talked through, we did eliminate multiple pieces of equipment simply because the customer opted not to use those pieces of equipment or because it just wasn't really amenable to their workflow. You also want to weigh the cost and reliability of an instrument accessory against the supposed convenience. And finally, you want to understand whether that instrument supplier will be a good long-term partner by offering good service and support. And so if you're only working with one instrument vendor, they of course will tell you that yes, they're a great partner. But again, by working with our field support scientists who have a lot of experience working with different automation vendors across the country, here in the US, and across the world, they really do know which instrument suppliers will be the most helpful to you in the long-run. So before we leave this scenario, I wanted to talk about the outcome for this customer. And of course as I mentioned, because this is modular automation, we're going to have a combination of manual and automated steps. For the manual steps, we actually left the customer with needing to prepare the cells and adding them to a reagent trough. They need to add the stock compounds in eppendorf tubes to the instrument. And then finally the user will actually prepare that dilution of the RealTime-Glo reagent and add the diluted reagent in a reagent trough. Once we load these things onto the instrument, the instrument can do the automated process, which will be to do the plating of the cells, preparing the serial dilutions of compounds, and we are actually preparing these in a 96 well plate and then adding the compounds from the 96 well plate into that 384 well plate containing the cells, and finally adding the RealTime-Glo reagent. Some additional lessons from working with this customer I wanted to talk about here on this slide. So when you're working with cells, or things that are in suspension versus solutions, it can be much more challenging. You want to avoid, um, so essentially you need to do a lot of mixing of the cells to make sure that you have a homogeneous suspension. And this is going to be critical for reproducibly plating cells, the same number of cells across the plate. For the same reason we recommend that you avoid aliquoting cells, or suspensions, by multidispensing, and when I talk about multidispensing in the HT world, I'm talking about aspirating kind of a bulk volume or enough total volume that you can dispense to multiple wells. And again, this is for the same reason that we need to mix. If you are dispensing through multidispensing, some of the cells are going to settle out as you dispense across the plate and potentially you'll get different numbers of cells plated across the plate. The next thing I wanted to talk about was to always optimize your process manually first. So initially this customer had done all of their pilot studies with control compounds and a 96 well plate. And so, the issue then is, so what we ran into then in miniaturizing to the 384 well plate is that we saw some differences in how those control compounds acted in the 96 well plate versus the 384 well plate. And so we weren't sure whether that was attributable to the automation itself or if it was an issue of scaling. So going back and doing that process manually in 384 well plates, as painful as that was, we found out that it indeed was the scaling factor. It did not have to do with the automation. And so we needed to optimize some of the conditions manually and then reimplement some of those changes into our automated workflow. The other thing I wanted to mention is that pipetting precision is much more challenging in these higher-density format plates because you're pipetting smaller volumes. So you do need to have a lot more experience with liquid handling. Because, for example, with precision, if you're trying to pipette ±10% at 10 microliters, that's a much harder target to hit than ±10% at 100 microliters. So you really need to have, if you are scaling down to these smaller formats such as 384 well plates or especially with 15,036 well plates, you need to make sure that you have somebody experienced in setting up those processes. And so we didn't actually just stop with the setting up of the assay plates with the biomech system. We did help this customer with more of an integrated process. So they actually had this biomech system like I said, we had it set up the plates, but then on one side of the instrument it had an automated incubator where we could incubate the cells and on the other side of the instrument was an automation-compatible plate reader. And so what we set the system up to do then was to move that assay plate that we prepared into the automated incubator. Sorry, I have some animations here that are being slow. And then the instrument using the kind of gripper robotic arm can move that plate into the cell incubator. And then at the different time points, so again, we're using the RealTime-Glo Assay that we can take multiple time readings, timed readings. We can use the instrument then to move the plate from the automated incubator into the plate reader. Sorry, my animations are being a little bit weird this morning. And so, with that then, that really unlocks the power of the RealTime-Glo reagent. Like I said, this assay reagent is used for measuring cell viability at multiple time points. So if you had to have an analyst come back and always be, like every four hours, for example, moving a plate from the incubator to the plate reader, that could be quite tedious again. And so if we automate that process, we really are able to further unlock the power of the RealTime-Glo reagent. So this kind of integrated system is ideal in this scenario. And so with that, I'm actually going to transition to the other scenario, which is using an automated liquid handler for a nucleic acid extraction, specifically DNA extraction. So this customer, in particular, was in a clinical diagnostic lab and they were interested in purifying genomic DNA from blood samples. This particular customer was processing thousands of samples per week and then using that purified DNA and various molecular analyses like qPCR and next generation sequencing. Because this is a clinical lab and they kind of need to keep track of chain of custody, there was a sample tracking requirement as part of this consultation. And so really what calls to automation here is the fact that they're processing thousands of samples per week. So that could take a lot of man hours from people working, like having analysts do this process mainly in the lab. And so the Promega tool that we used for the automation here is the Maxwell HT Genomic DNA Kit. This kit provides all of the reagents that you need for DNA purification for multiple sample types, including blood. And it's based on the use of paramagnetic particles that react in a magnetic field. These paramagnetic particles bind the DNA. So we can bind the DNA on these paramagnetic particles and then carry it through a series of washes before finally eluding the purified DNA. We're pairing this Promega tool with the Hamilton Mag X Star in this case. And in this scenario, the customer actually hadn't selected a platform that they wanted to use. They actually worked with our field support scientists identify the platform that was going to work best for them. And this Hamilton Mag X Star is actually a system that is purpose-built for DNA or RNA purification using paramagnetic particles. So it's already kind of set up with all the equipment that they're going to need for their processes. So the first thing in breaking down the workflow is that we need to reformat the samples. And this is a place where we're going to use those individual channels, because we're going from less automation friendly blood tubes. So these are 16 millimeter blood tubes that the customer was collecting in. We need to transfer out of those not so automation friendly tubes into an automation friendly 96 well deep well plate. And so that is going to be the first step of the protocol is to transfer the samples into the plate. Once we have all the samples in the plate, we're going to treat everything in the plate the same way and we're going to carry it through this nucleic extraction workflow. Now, I'm not going to walk you through this. This would be really tedious to talk through. And so just to kind of give you a summary of the basic steps of nucleic acid extraction, first you need to lyse your cells to release the nucleic acid to the media. And then we are going to bind that nucleic acid on those paramagnetic particles. We'll collect those paramagnetic particles after we've done the bind, and remove the waste. Then we can start our wash processes where we add and remove wash buffers from those paramagnetic particles. And finally then we can elute in just aqueous buffer at the end of the nucleic acid extraction protocol. And so if we look at what kinds of processes we need to incorporate here, there are just a few things that kind of indicates what pieces of equipment we will need to use here. So obviously we're going to need the ability to pipette because we're aspirating and dispensing reagents. We do use heat during our lysis protocol, so we need some way of heating the sample. We also need a means of moving the plate. So we're going to be moving our samples on and off the heater and on and off a magnet to manipulate those paramagnetic particles. And then we also need to be able to shake. So in terms of pipetting, again, we are going to use both means of pipetting, both individual channels and the 96 well heads. As I've kind of reiterated at the beginning that we want to use the channels where we're transferring from those individual blood tubes into the processing plate. And we're also going to use the channels at steps of the protocol where we're using smaller volumes of reagent, where we need to use lower, need to have lower dead volumes because there's not like several milliliters of overage of those reagents. We will use the channels to handle those reagents. We will also use the 96 head as well. And the 96 well head is used to really speed up the process of processing these samples. We will use the 96 head to transfer bulk reagents, such as the wash buffers, where we do have a lot of volume that we can play with, and removing wash buffers and stuff from the samples and the processing plate, and we'll also use it to transfer the eluent to the final elution plate. The next piece of equipment that we'll need is one that we can use for heating and shaking our samples. And it's very common on these systems to find a combination heater-shaker device. I just wanted to again note some words of advice here about choosing what type of device you actually want to implement. For this protocol, again, we're working with the paramagnetic particles. We want to keep them in suspension. And so we need to think about how we are going to be shaking and what's going to be the most effective way to keep those magnetic particles in suspension. For the shaker devices, you can either get linear or orbital shake patterns. So linear shakes would just kind of go back and forth, whereas an orbital, you're kind of going around in circles. And you can have two or three millimeter orbits. And so to get the best mixing here for this protocol we use a shaker that has an orbital shake pattern with a 3 millimeter orbit, because that is just going to get more kind of mixing motion in the wells. The next thing has to do with the heating capabilities of these devices. So as you can see on this device from Hamilton, the heat plate is just a flat plate across the bottom. And if you just use that kind of flat plate and heat it, you'd get very inefficient heat transfer. So they do produce heat, specific heat adapters, that are compatible with different types of plates that you might be using in your process, and those basically cut the bottoms of the wells to improve the heat transfer. However, heat transfer is never perfect, so we do tend to set the temperature on the device higher than the intended temperature of the sample. And finally, I wanted to mention, like your oven at home, preheating is required. It's not as simple as, it's not like a thermal cycler where it will ramp up the temperature extremely quickly. You do need to give it time to get to what set temperature you want to process your samples at. And next, like I mentioned, we're going to need to be able to move the plate on and off that shaker and then on and off the magnet. And there are a couple of different ways that we can do that. We can use a plate moving arm or we can use gripper paddles. The plate moving arm is a more expensive accessory, but it is more flexible. So if you have steps in your process where you might need to reorient your plate from like landscape to portrait format, it would be appropriate to use one of these plate moving arms. So you can see in the picture here that we have these kind of gripper fingers that come off of a motor mechanism that allows you to kind of manipulate the plate in 360 degrees. So if you did need to reorient your plate, you would need to use plate moving arm device. If we are moving plates outside of the instrument, that would also be a place to use these plate moving arms For the gripper paddles though, these are fairly inexpensive accessories, and on the Hamilton system they use these gripper paddles that actually just attach to the pipetting channels on the instrument. So basically they just squeeze onto the plate to pick up and move it around. But because we are just kind of basically squeezing the plate between the two gripper, or the two pipetting channels, we can't reorient the plates and we can't use it to reach outside of the instrument. However, for this particular customer who is interested in DNA extraction, we don't need to reorient the plate and we don't need to move it outside of the instrument. So these gripper paddles are really more than sufficient for what we need in this process. So we opted for the gripper paddles for this particular customer. Finally, as I had mentioned early on in the scenario, the customer does have a sample tracking requirement. And so the Hamilton system that they ended up with uses these tube carriers. And these tube carriers are specific to the type of tubes, these 16 millimeter blood tubes that they're using. And so what you can see on the right is the barcode reader. So as the instrument slides these tube carriers into the instrument, the barcode reader will read the barcode on the sample tubes as well as their position in the tube rack. And that helps us keep track of where the samples are being processed within the plate. So the final outcome for this customer, again, I want to emphasize that with modular automation, we're looking at some manual steps plus automated steps. The user is going to load the uncapped blood tubes into the tube carrier. Then load the reagents that we're using for the purification into the appropriate reagent troughs on the instrument. And then they're going to add the consumables to the instrument. So these are things like the tips and processing plates. And depending on how many samples they're studying in the process, this will take about 30 minutes of hands-on time to set up the instruments. Then once we have the system loaded up with our samples and our reagent, the instrument's going to take over and dispense the blood into the deep well processing plate. And then this method actually that we set up for the customer will process up to 384 samples in about two hours. So we're able to process 4 96 well plates in parallel. So if you think about how long that might take you, if you were trying to do that manually, that would be very slow. That would probably be more than or it would be several working days for one person to do. But we can accomplish that in about 2 1/2 to 3 hours total between the manual and automated steps. So with the couple of scenarios that we've talked through today, we've discussed some of the questions to ask when we're considering automation. This first question, of course, we chose a couple of processes that would allow for automation. So the cell culture analysis and the DNA purification. We also described what processes did we actually want to automate to save time, and what exactly we wanted the automation to achieve. Some of the other things that you want to keep in mind are things around what are your quality requirements? So for the DNA purification customer they might have specific yield requirements or purity requirements that we can help them to achieve with our chemistry on the automated system. When is the project deadline? So automation projects never run quickly. I would say probably you need three to four months at the very minimum to get a project going. There's always a lag time for actually receiving an instrument once you've decided on one. And then you'll need to develop the method, put that on the system and validate the method as well. So three to four months might even be short, but these projects do tend to take some time, so plan for that when you're implementing automation. You want to think about where the automated workflow will be done. Of course, the instruments themselves do require space, but there may also be particular electrical requirements that need to be accommodated for these systems. You need to think about who in your group will operate and maintain the workflow and instruments. I really strongly recommend that you have multiple people on this so that knowledge doesn't leave with a person who might move to another lab. And finally you want to think about what is your budget? And that's, of course money is always a question in these scenarios, and in considering your budget, it's going to determine what actual automated system you can get. So in summary, I want you to have taken away a few things from this talk. The first thing is that you want to understand your process at the most fundamental level by mapping it out. You'll prioritize automating the parts of the process that will lead to the biggest time savings. Determine what instrument accessories are required to support your workflow automation. And then the last thing I want to challenge you to think about is to consider opportunities for expansion into other processes. So you might have one particular problem in your lab that you're considering adopting automation to help resolve. But pieces of automation equipment can be used for multiple processes. So if you can see, kind of into the future, that you might want to expand automation for other lab processes, think about equipping your instrument in a way that will make it most flexible to use across multiple different processes in your lab. And with that, I wanted to ask a question here. So when do you plan to implement automation in your lab? Are you looking to do this in less than six months, the first option? Less than a year? Greater than a year? Or are you just looking to learn a little bit more about automation right now? And so you guys can go ahead and click one of the options here and we'll take a look at the results here in a minute. And I see some answers trickling in here. Thank you guys. Got some going here. I'm going to move, you guys can keep answering, I'm going to move on to the next slide here. Most of you it looks like are just looking to learn a bit more about automation. That's a totally valid place to be in your process here. I think that that's, I mean, you need to start thinking about it somewhere and just starting with some basic education about automated systems is definitely very helpful. All right. And so with that, I just wanted to touch on, again, the support that Promega can provide to customers. And so our field support scientists can help. And kind of the first step of that process is to choose the Promega assay or kit that you want to automate. And then once you've got that kit, you'll connect with the FSS team either through an automation inquiry form, which we have linked in the Resource Center of this webinar, or you can connect with them through your Promega sales representative. Then they'll consult with you about what processes you're looking to automate and what exactly do you need. So they'll talk about things like what do you need the instrument specifically to do? Are there specific technical features that will help to support your goals? Again, they are a really good resource for information about whether a particular instrument provider will be a good partner. They have these insider knowledge about, because they work with so many customers across the world, they know in particular regions of the world, one instrument supplier might be a better service provider than another. These are all things that will factor into what particular instrument you want to choose. And finally, and this has been coming up more kind of as COVID-19 testing has scaled down from what it was during the initial years of the pandemic, people adopted automation during COVID-19 to do COVID-19 testing, and so they might not be using that instrumentation for that testing anymore. So can we retrofit and use those instruments that had been used for the COVID testing into a new application? So they will consult with you, you'll place an order for the instrument that you guys kind of agreed upon will be the best for you. The Promega FSS team actually writes the script. They'll install and optimize the method with you on-site and they'll work with your staff to train them. They usually spend about a week on-site depending on the complexity of the project that they're working on. And then we can offer continued support, so it's not just one and done, we've installed it, we're hands off. You can always contact the FSS through technical service if you do run into issues with your automated method. So again, I just wanted to mention how you can reach out to the field support scientists. Again, you can connect through your Promega sales representative or you can connect through the Automation Inquiry form on Promega's Laboratory Automation resource page or it's also available in the Resource Center within this webinar platform. And so with that, I wanted to introduce Brandon Krieger. He's actually one of the automation scientists on our field support scientist team. He's been with Promega for several years now, but has a lot of automation experience from his background with other companies. And so I brought him on. So if you have specific kind of instrumentation questions, in that you need that insider knowledge, Brandon's here to help answer those questions. And so with that, I'm going to open the floor up to some questions. Thank you, Sarah very much for a really good presentation. We do have some questions that have come in. So I'll start us off with one from Mohammed. He's asking about your discussion of the, the blood extraction, and he's asking what about the the risk of cross-contamination when the instrument will transfer the blood sample from the initial sample tube into the plate. So kind of how do you consider that? And when we develop methods, how do we demonstrate that we're achieving a method that doesn't have cross contamination? Yeah. Thanks, Eric. Hi, everybody. My name is Brandon Krieger, as Sarah had mentioned. You know, there are different options depending on the instrument that you are utilizing. One of the things that we will typically go through and do, is kind of create safe paths, safe paths for the channels or pipetters to travel in transferring to that plate. So there are many ways that we can minimize cross-contamination. As well as, you know, depending on the instruments, technology and capabilities. You know obviously one of the things that you can run into with blood samples are clots in those associated blood samples. So there are different ways to mitigate that and we've gone to the extent in the past of even programming kind of a, you know, essentially a tip drag across the top of the sample tube to ensure that if there are any blood clots that are hanging from the end of the tip in order to prevent cross-contamination. You know, essentially just create a tip drag across the top to clear any clots that may be in the way. And then we can create safe travel to the plate to minimize cross-contamination. And one of the things that we frequently do, as well, in that process, is through a lot of our testing we'll do somewhat of a checkerboard pattern just to ensure that we're not really seeing cross-contamination through the process. Yeah, specifically with blood, so I mean, what I do as an application scientist is kind of a lot of the behind the scenes work for these methods. And so one of the things that we will do, like Brandon said, is a cross-contamination type of analysis with a checkerboard of samples. So we'll checkerboard male and female blood essentially. And then because male blood has that Y chromosome in it, we'll look for contamination with the Y chromosome in any of those eluates that were extracted from female samples. And we have very sensitive ways of detecting that Y chromosome. And so that's definitely one way that we can check to make sure that we don't get cross-contamination. All right. Thank you guys. Another question from Julia. And Julia is asking, do you have special hints for automating DNA extraction from buccal swabs? And she was actually I think specifically asking on a Hamilton. Yeah, so buccal swabs, you know, obviously can be a challenge for a liquid handler in general. I know that there are options within the field, especially Promega themselves have what they call the MaxSpin 96, which is basically a basket that can be utilized in a 96 well format. So you clip the the buccal swab heads off into the MaxSpin basket and it allows you to add the lysing solutions to the, you know, that basket that is nested within a 96 well plate and there is a, you know, essentially perforations in the bottom of that basket, which allows you to complete the lysis in the presence of solution. And then what you can do is you then separate that basket from the 96 well plate through a centrification. So there is some manual manipulation that is required during that centrification step. However, as Sarah had mentioned, you know some instruments can be, you know, integrated with centrifuge instruments as well. So there is the potential of automating that in some format. But, you know, what it does, is it helps in that initial stages of creating a lysate that is then separated from the buccal swab itself. Okay. Next question from Sean. Sean missed the beginning and is asking do we provide different liquid handling solutions aside from Hamilton? And then you also you mentioned validation, do you have experience with your liquid handling solutions under the GMP environment? As far as liquid handlers, Sean, I can quickly answer that, really it was just an example that Sarah provided that happened to use Hamilton's. We've got experience and it worked with really almost all the major liquid handling and particle movers that are sold worldwide. So we support our chemistries pretty much on any automation system. As far as validation, I'm going to turn that probably over to Brandon to see what what you might say as far as kind of how we might support there. Yes. So from the validation side, what we would do is we would set you up with the appropriate method. You know, I understand coming from a CRO in my past life, that validation under a GMP setting and environment is challenging. What I can say is that our reagent manufacturing is completed under ISO requirements. So that lends a benefit to the validation on your end. But basically the validation would just need to to follow your your specific SOPs within your own laboratory for, you know, for the validation end of things. Make sure that we're covered. Okay, we're approaching time. There's one more that we'll try, I think. And we may answer this one more offline. And that's do you have a kit for RNA extraction of tissue that was stored in formaldehyde only and not FFPE. I've tried out RNA FFPE kits and it didn't work well. Do you have recommendations for kits for processing these types of samples? That may be a very, a very challenging sample type. Sarah, do you know in your experience, have you had much experience with, it would be highly crosslinked samples because they're essentially stored in the fixative. So they're going to be highly crosslinked. Right. These will be particularly challenging samples. We have worked with things like, so one of the actual cervical sample collection devices here in the US, SurePath devices do contain formaldehyde. And so we have worked with, like cells from those devices, and basically like Eric said, I think it'll be best to kind of take this offline for more communication. I do have suggestions, I don't know that the FFPE kit will be the best option here because it's designed for smaller amounts of sample. I don't know how large the tissues are that you're working with, but we might want to use more of a tissue-specific purification chemistry versus FFPE to be able to capture a lot of the nucleic acid and then incorporate steps to de-crosslink the sample, the nucleic acid in the sample, to the best extent possible. But like Eric said, it it will definitely be a challenging sample type, and it's one that we could, that's kind of my side of things where we get to work with all of these sample types that you guys bring in as particularly challenging. We develop those methods initially and then we can kind of hand those off to the FSS for implementation on automated platforms at customer sites. So yeah, thank you. We'll be in touch. Well, everyone, we, oh go ahead. Yeah. Eric, there is actually one more question that did pop up earlier, and it was regarding a sterile environment in a BIOMEX system. And there's a couple of options that people typically go with. One, they will try and put these instruments into a sterile room. The other option is, is I do believe most of these instruments, including the BIOMEX, do have an add-on for like a HEPA filter hood, to kind of create and make it a biosafety cabinet itself. So I just wanted to make sure that that got out there too. Yeah. And for this particular customer, they weren't concerned with sterility, essentially they were plating the cells with antibiotic and I realized that's not ideal for some cases. So we didn't take those kind of sterile measures or they didn't in their lab, but they're, like Brandon said, there are possibilities to do that on the different platforms. Okay. Well, thank you, Sarah. Thank you, Brandon. And thank all of you for joining us today. We've reached the end of our time together. If you have any other questions, please feel free to reach out. We'll also reply with written answers to the questions through the Q&A as well to those of you that have asked. But thanks again and thank you for joining us. We do have a short poll that will show up at the very end here and we really appreciate your feedback and input on our webinar series so that we can continue to provide content like today that is engaging and meets the needs that you have in learning more about Promega's products or our activities in automation and science. So thank you very much and have a great day everyone. Bye. _1732399961158