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• Patricia Atkins Senior Applications Scientist at Spex® CertiPrep
• Alan H. Katz, PhD Director, Global Product Manager – Chemicals at Spex CertiPrep
• Eric Smith Applications Scientist at Spex® SamplePrep
• Lea Anderson-Smith, PhD o Director, Engineering (R&D) at Spex SamplePrep
Polymerase Chain Reaction (PCR) is an essential tool for biological analysis. This molecular biology technique gained newfound recognition when it was thrust into mainstream attention during the onset of the COVID-19 global pandemic; and PCR testing became the standard tool for detection of the disease-causing virus (SARS-CoV-2).
Since its invention in the 1980s, PCR has become instrumental for many applications such as medical testing, biological research, quality control, forensic analysis, and environmental monitoring.
PCR technology utilizes the biological DNA replication machinery to amplify small quantities of genetic material to facilitate downstream analysis—such as microbiological detection and genetic fingerprinting. Therefore, it is important to carefully process and prepare PCR samples to minimize contamination and increase available material.
Equally important is understanding all the key steps, interactions and chemical reactions that take place during the amplification of genetic material to generate usable and quantifiable results. In this presentation, we will look at the basics of PCR and the practical aspects of sample preparation, chemical reagent selection, and types of processing that can improve or amplify the PCR workflow.
Despite its mistaken reputation as a difficult technique to master, PCR is, in fact, a simple tool that offers accurate and rapid results. Whether you are involved in clinical diagnostics, medical research, bioprocessing, vaccine and therapeutics development, food and beverage, agriculture, or botanicals testing, you can use PCR to deliver quick and consistent results.
Antylia Scientific™ (formerly known as Cole-Parmer) and Spex® have a range of innovative tools that can be leveraged for rapid, reliable, and reproducible PCR analysis.
Patricia Atkins — Senior Applications Scientist at SPEX® CertiPrep
Patricia graduated from Rutgers University in NJ and started her career as a laboratory supervisor for Ciba Specialty Chemicals. In 2008, Patricia joined SPEX CertiPrep within their Certified Reference Material’s division. As a senior application scientist, she spends much of her time researching industry trends and developing new reference materials. She is an experienced speaker and has presented at numerous conferences, including NACRW, NEMC, Pittcon and AOAC. She is also a published author and her work has appeared in various journals and trade publications, including Spectroscopy, LCGC and Cannabis Science and Technology, where she is a columnist for analytical issues in botanicals testing.
Alan Katz, PhD — Director, Global Product Manager - Chemicals
Alan holds a BA in Chemistry from the University at Buffalo and a PhD from Princeton University. His specialties include organic, computational, heterocyclic, and synthetic chemistry, drug discovery, biological assay design, structure-based molecular modeling, and lab automation. He has enjoyed combining his expertise with his interest in computer programming to develop a system to assist in drug discovery. At SPEX CertiPrep, he oversees the manufacturing and quality control of certified chemical reference materials, including inorganic and organic chemical standards, NPD and R&D.
Eric Smith – Applications Scientist at Spex SamplePrep
Eric holds a BS in chemistry from Kean University. His current role at Spex SamplePrep involves managing all phases of application development including specifications, data collection, literature research, reporting and presentations. He serves as applications scientist and an internal ISO 9001 auditor; a technical expert, traveling domestically and internationally for customer visits and tradeshows. Prior to joining Spex, Eric worked as a R&D and QA chemist and currently also serves as an instructor at the International Center for Diffraction Data and the London X-ray Consulting Group.
Lea Anderson-Smith, PhD – Director, Engineering (R&D) at Spex SamplePrep
Lea Anderson-Smith holds a BS in Chemistry from the University of Wisconsin and a PhD in Inorganic Chemistry from MIT. She is currently Director of Engineering for Spex SamplePrep and Katanax equipment and accessories, managing design and development of new products and updates to existing products. She also managed operations for the Spex SamplePrep business for many years and maintains an active role in technical support for customers and service of customer-owned equipment. She works closely with the product management and commercial teams in identifying new product and application opportunities with the goal of helping scientists in their work.
Madalina Enache – Product Sales Manager (EMEA) – PCRmax®, Techne®, and Argos Technologies®
Madalina holds a BSc (with Hons) in Biology and a MSc in Microbiology. She joined Antylia Scientific™ three years ago, when it was known as Cole-Parmer, as part of the application sales team. She has risen through several commercial roles, focused on PCR products, to become the EMEA Product Sales Manager for our PCRmax, Techne, and Argos Technologies brands.
Lysing of Bacterial Cells in the Geno/Grinder
With kind permission of Russell H. Vreeland, West Chester University, January 2003
The Geno/Grinder was tested to determine if this technology could be used to lyse bacterial cells. Standard 96-well titer plates were used with 400 to 600 μl silica grinding beads (Molecular Biology Grade, cat. no. 2166).The delivery of the beads into each cell of the titer plate can be accomplished in a number of ways. In this case micropipette tips were filled to the mark with grinding beads, and each tip emptied into a titer plate well. This technique will deliver approximately 0.4 grams of silica beads per well. Other delivery systems are commercially available. Experiments were performed with 250 to 500 ml of bacteria: in one case the gram-negative, salt tolerant Halomonas elongata, and in the other, gram-positive Bacillus. Both organisms were grown to late log state at 37°C. with shaking, then harvested via centrifugation at 7000 RPM for 10 minutes. 0.4 grams of silica beads were added to each well, and the plate was sealed. The filled titer plates were then shaken in the Geno/Grinder at a setting of 1450 strokes per minute, in one-minute intervals for periods ranging from one minute to twelve minutes.Culture filtrate from the shaken plates was then measured for optical density at 260 nm. The average data for three separate “well sets’ is shown in Figure1.
The Geno/Grinder is capable of lysing bacterial cells in 96-well titer plates with 400 to 600 μl silica beads as grinding media. With increased grinding time culture filtrate shows a definite increase in optical density, indicating release of nucleic acids during grinding.Grinding times of 6 to 9 minutes appear to be suitable for producing measurable amounts of nucleic acid.