Simplifying HT Nucleic Acid Purification. With Expert Automation Support

Transcription

1 Simplifying HT Nucleic Acid Purification With Expert Automation Support

2 Housekeeping Application Widgets Movable and resizable Automation Experts ready to answer your questions Use Q&A widget now through 15 minutes after the presentation Resource materials available Use Resource List widget Download resources Bookmark links Please complete the survey

3 Optimizing Nucleic Acid Purification Workflows and Downstream Assays

4 Purification Workflow Lyse Cells Bind Nucleic acid Wash Impurities Elute DNA or RNA 4

5 Which Purification Scale is Right for You? Manual Column Single Samples Small Automated Maxwell 1-16 Samples 96-well Manual/Automated 96-well Vacuum-based Bead-based Automated 96-well Silica or Cellulose Magnetic-based 5

6 What roadblocks are you experiencing when automating your purification workflow? A. I m overwhelmed B. Sample challenges C. No time D. Robots are complicated 6

7 Common Barriers to Automating Purification Struggling to get desired results Challenging sample types Lack of time Complexity of robotic instrumentation 7

8 Expert Automation Support Scientific Experts in Purification at Any Scale

9 Supporting HT Applications Dedicated Teams Field Field Support Support Scientists Scientists Scientific Field Application Applications Scientists Integrated Application Solutions Scientists & Engineering R&D Scientists Technical Services Services Scientists Instrument Experience Tailored Reagents 9

10 Benefits of Customized HT Purification Support Make sure your instrument is working properly Create a customized purification protocol Decrease implementation time Avoid Common HT Purification Difficulties Don t have to learn complicated programming language Don t have to spend time optimizing instrument methods Don t need to troubleshoot chemistry issues that may/may not be related to instrument 10

11 Avoid the stress of.. Inability to scale up purification, keep up with demand Expensive instrumentation that goes unused Unreliable downstream results

12 What is the most stressful part of lab automation for you? A. Instrument reliability B. Difficult samples C. Programming D. Chemistry performance E. Money F. Time G. Overwhelming options H. Finding help you can trust 12

13 Supporting HT Applications Dedicated Teams Instrument Experience Capabilities include most liquid handlers and particle movers Tailored Reagents 13

14 Support for ALL Your Research Needs Consultation and assistance for troubleshooting and improving laboratory efficiency Development and optimization of automated methods to meet your needs Any scale Any sample type Any instrument Expertise to support your workflow from assay design to end point measurement Provide direction and identify appropriate solution Analyze data to systematically investigate and correct issues during development Customized training associated with new protocols to ensure your staff are confident and ready

15 Supporting HT Applications Dedicated Teams Instrument Experience Tailored Reagents Maxwell and manual chemistries available in HT kit format Easily customized 15

16 Customer Story: DNA & RNA Extraction from the Same Sample Problem: Needed an automated process to extract both gdna and RNA from same lysate in 96-well format Solution: Developed a protocol that purified RNA in one workflow and DNA in another workflow from same starting lysate Worked with customer to determine: Extracted DNA & RNA requirements Workflow needs Current automation configuration On-site visit: Used Maxwell 16 Instrument as POC to show chemistry was effective 16

17 Customer Story: DNA & RNA Extraction from the Same Sample From Test to Full Customized Automation Onsite testing performed on same day Maxwell 16 simplyrna Cells Kit vs. simplyrna Automated on Beckman FX K562 cells per extraction on both platforms Three samples processed with Maxwell 16 Instrument and Kit Beckman FX Test Used simplyrna bulk reagents (same Maxwell chemistry) Processed 96 samples in 96-well plate Miniaturized the extraction volumes so individual samples could be processed in each well of the plate 17

18 Customer Story High Throughput Plasmid Purification from 30ml Bacterial Cultures Problem: Needed an automated process for plasmid preps on a Tecan EVO system as part of a workflow to process many samples and develop therapeutic antibodies. Plasmid purification needed to be at a midiprep scale in order to effectively transfect cells. Solution: Developed a protocol that could: Process plasmid preps from 30 ml of bacterial culture Be automated on a Tecan EVO system Return greater than 100 µg of plasmid After testing plasmid in the workflow, all of the criteria were met and the solution became part of the workflow. 18

19 Customer Story High Throughput Plasmid Purification from 30ml Bacterial Cultures Average yield and purity of plasmid DNA isolated using the automated Wizard MagneSil Tfx System from 1.5ml of bacterial culture of JM109 transformed with pgl4.50. Panel A. Yield and purity of plasmid DNA were measured by absorbance using a NanoDrop One Spectrophotometer. Panel B. Yield was measured by fluorescence with the QuantiFluor ONE dsdna System on a GloMax Discover Instrument. Culture 1, N=32; Culture 2, N=32; Culture 3, N=30. 19

20 Customer Story High Throughput Plasmid Purification from 30ml Bacterial Cultures Yield and purity of plasmid DNA isolated from 1.5ml of bacterial culture using the automated Wizard MagneSil Tfx System Yield (Panel A) and purity ratios (Panels B and C) of plasmid DNA were measured by absorbance using a NanoDrop One Spectrophotometer. Averages were calculated for the full plate and are indicated by lines on the graphs 20

21 Customer Story Purify Total Nucleic Acid (TNA) from Fecal-Derived Bacterial Cultures Problem: Needed an automated process for the purification of total nucleic acid from Gram positive and Gram negative bacteria Bacillus, E.coli, Clostridium, Methicillin-Resistant Staphylococcus aureus (MRSA) and Salmonella on a Tecan Fluent and KingFisher Presto system. Solution: Developed a protocol that could: Process bacterial pellets from overnight bacterial cultures Successfully lyse and release DNA from both Gram positive and negative cultures Be automated on a Tecan Fluent and KingFisher Presto system KingFisher Flex was used as POC to provide preliminary quantitation data on the purified TNA eluates. KingFisher Presto was a newly released product and, therefore, not available for testing. 21

22 Customer Story Purify Total Nucleic Acid (TNA) from Fecal-Derived Bacterial Cultures Bacterial TNA (dsdna) eluate quantitation on the NanoDrop ONE spectrophotometer 22

23 Customer Story Purify Total Nucleic Acid (TNA) from Fecal-Derived Bacterial Cultures dsdna purity ratios for bacterial TNA eluates on the NanoDrop ONE spectrophotometer 23

24 Which is your greatest automation challenge right now? A. gdna purification B. ccfdna purification C. FFPE purification D. Plasmid Preps E. PCR/NGS Cleanup F. Quantitation/Normalization G. Reaction Setup 24

25 How Custom HT Purification Works

26 Automation Support Made Easy Request a consult Create a customized plan With guidance and support at every step from: Field Support Scientists Field Application Scientists Application Scientists R&D Scientists Technical Services Scientists On-site implementation

27 You could win our Maxwell HT VIP Customer Experience: All expenses paid trip to Promega headquarters in Madison, WI Meet with R&D Scientists and Maxwell HT Support Team Featured in an upcoming webinar about your experience To enter, simply Request a Consult for help with your purification questions. 27

28 Thank you!