http://www.rssboard.org/rss-specification 720 https://escholarship.org/uc/ucdcrf/rss Recent eScholarship items from Contained Research Facility Fri, 19 Jun 2026 10:27:46 +0000 https://escholarship.org/uc/item/6cb1956t <p>This manual outlines a molecular method of identifying tomato leafminer without the need for specialized equipment. The method is based on the CRISPR-Cas (Clustered Regularly-Interspaced Short Palindromic Repeats – CRISPR associated proteins) system. In this system, the Cas12a enzyme has indiscriminate single-stranded DNA nuclease activity when it binds to target DNA. If a single-stranded oligonucleotide probe that has a fluorophore and quencher is present when the Cas12a enzyme binds to the target DNA, the single-stranded oligonucleotide probe will also be cleaved, releasing the fluorophore and quencher.</p><p>The fluorescent signal which is proportional to the number of target DNA molecules, can be measured. The Cas12a enzyme reaction is isothermal and occurs between 15°C and 50°C. Coupling this reaction with Recombinase Polymerase Amplification (RPA), an isothermal DNA amplification method, the target DNA from the tomato leafminer can be detected and visualized using a fluorimeter... https://escholarship.org/uc/item/6cb1956t Tue, 18 Mar 2025 00:00:00 +0000 Tabuloc, Christine Lewald, Kyle M Eweis-LaBolle, Daniel Song, Wenqi Godfrey, Kris Chiu, Joanna