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22 | MAY 1, 2015 | GENengnews.com | Genetic Engineering & Biotechnology News Genotyping Using CGE OMICS A Novel Single-Channel Microfuidic Device for Nucleic Acid Fragment Analysis Varoujan D. Amirkhanian, Shou-Kuan (Eric) Tsai, Zsolt Ronai M.D., Ph.D., and Gene W. Stewart Nucleic acid analysis, genotyping, PCR, and RFLP methodologies are time-consuming techniques using agarose or acrylamide gel electrophoresis—image capture plus gel im- age interpretations. Capillary gel electropho- resis (CGE) automates these, providing fast separations with high sensitivity, excellent res- olution, and ease of operation. However, de- spite its effectiveness current CGE has a high cost of instrumentation and cost per sample. Much of this cost is due to traditional designs. BiOptics' Qsep100™ CGE system solves this problem by integrating microcapillary electrophoretic technology into a plug-and- play disposable pen-shaped capillary gel cartridge with automated liquid handling, real time fuorescent detection, and analy- sis. Optimized for high resolution and high throughput at ambient temperatures, the need for expensive, extreme voltage power supplies and external cooling is eliminated, thus reducing instrument costs and increas- ing productivity. Genotyping and other relevant applica- tions with sample throughputs to 96 can be automatically analyzed within 5–6 hours. By integrating and automating all analytical functions required for the various types of nucleic acid analysis, Qsep100 provides im- proved peak resolution, good linear dynamic range, and reproducible migration times with a lowered cost. System Design Overview Qsep100 was designed to integrate, au- tomate, and miniaturize the laborious tech- niques of traditional DNA electrophoresis, and reduce the costs associated with tradi- tional CGE. The Qsep100 CGE system (Figure 1) is composed of the core instrument (38 x 39 x 40 cm), an automated liquid/sample handling (X-Y-Z) platform, real-time fuorescence de- tector, and multi-use disposable pen-shaped capillary gel-cartridges; all are driven by an intuitive GUI with analytics. The platform contains buffer(s), a sample waste reservoir, DNA ladder(s), and samples arrayed in a 96- well format. This design provides fexibility in single-channel CGE instrument applica- tions at low cost-per-sample run analysis of DNA fragments. The unique pen-shaped capillary gel-car- tridge (CGC) permits plug-and-play use in a robust, injection-molded body with an inte- grated gel-ethidium bromide reservoir and a short length single microfuidic glass capil- lary column (75 µm ID, 15 cm total length, 11 cm separation length). The shortened capillary length allows for reduced operating voltages (1–15 KV), shorter run times, and the elimination of expensive cooling systems such as Peltier or recirculating chillers. To mitigate electroos- motic fow (EO) and prevent analyte adhe- sion to the inside walls of the capillary col- umn, they are dynamically coated with the separation matrix. This design also includes top and bottom electrodes (anode and cathode), a detection window, imbedded radio-frequency identif- cation (RFID) chip, and a label to identify the gel-cartridge and monitor its number of electrophoretic runs. Each cartridge contains a proprietary gel/ethidium bromide mixture and is capa- ble of analyzing 200 samples in 2 minutes per sample analysis, consuming as little as 1 pl from sample volumes between 1 µl to 20 µl. Real-time sample detection is achieved by an innovative optical fber system (Figure 2) Figure 3. (A) Installation of the capillary gel-cartridge. (B) Bufer solution and ladder tray with 96-well sample tray. A B Figure 2. (A) Photograph of the ball-end dual fber optic system. (B) Schematic of the LED- induced fuorescence detection system with pneumatically operated fork assembly, which aligns two optical fbers precisely to the detection zone of the glass capillary by clamping to the two pre-installed plastic ferrules (excitation @ 530 nm; emission detection @ 590–620 nm). A B Figure 1. (A) BiOptic's Qsep100. (B) The Q-Analyzer software with easy-to-use GUI for easy operation and rapid data analysis. (C) Multi- use disposable pen-shaped capillary gel cartridge. A B C Tutorial Contact us today 1.800.523.0072 www.custombiogenics.com/GEN Custom Systems BioGenic Chest Racks LN2 Racks Mechanical Freezer Racks Bag Canisters & Frames Custom Racks Liquid Nitrogen Freezers Cryosystems & Dewars Lab Tanks Remo te Monito ring Systems Better Product Reproducibility Documented Results Custom Systems BioGenic Your Complete Solution Inventory Systems Controlled Rate Freezers Long Term Storage & Transport Monitoring Custom BioGenic Systems ofers a complete solution for freezing and storing cell lines and biologicals