- Ironwood Ridge High School
- The Ridgewood Methods
Manno, Theodore
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THE RIDGEWOOD METHODS
FROM GENE TO PROTEIN
DNA EXTRACTION, PCR, ELECTROPHORESIS, CLEANUP & ANALYSIS
The Ridgewood Methods are composited, modified protocols for complex laboratory procedures intended specifically for high school instruction at IRHS. They are easier to understand, more straightforward to implement, and more operable within 60-minute high school class periods than some other protocols. Students learn these techniques as part of their broadly-based training in high school, so as to provide the background skills to pursue a speciality later in their careers.
Because of the COVID-19 pandemic, once-specialized vocabulary (e.g., neutralizing antibodies, PCR, mRNA) is now part of the everyday lexicon. Using safe, inexpensive, yet powerful models for these concepts (such as a rubisco protein subunit in leaves from campus plants) is not necessarily straightforward, but facilities and training at IRHS are modernized and cutting edge. As an additional focus, we also learn about conducting the different types of Polymerase Chain Reaction (PCR), primarily as it relates to (simulated) COVID-19 testing and forensic applications.
We sometimes make even further adjustments to these protocols when time is of the essence during a shortened period or the day before a weekend or holiday. In haste, PCR product is sometimes cleaned by centrifugation with "Manno's Buffer"--80 mL 95% EtOH mixed with 20 mL 0.3 M NaOAc, 1 tbsp salt, and 1 tbsp of meat tenderizer.
It's an exciting time to be a Nighthawk. Join us at Nighthawk Biotechnology-Next Generation. We have awesome lab equipment, and awesome stickers. Chowl stickers.
Sources: (1) DNA Learning Center (DNALC) CSHL DNA Learning Center (2) Fractionation of DNA Fragments by Polyethylene Glycol Induced Precipitation by John T. Lis in Methods in Enzymology, 65 (1980), pg 347-353 (3) Purification of PCR Products for Sequencing http://www.biology.ualberta.ca/facilities/MBSU (4) Instructional Manual for A Process to Dye For, lab exercise from Flinn Scientific.