Real-Time Isothermal DNA Amplification with Quantitative Answers in 20 Minutes or Less

Advantages of the real-time Nexar system include:

• Fast, accurate, real-time results in 20 minutes or less for a 384-well array.
• Fully automated assay setup and detection system enables scalability to high throughput.
• Flexibility to run low volume (1.6uL) isothermal and qPCR chemistries on the same instrument solution.

Instrumentation

The Nexar Optimized for Real Time Nucleic Acid Processing is a fully automated solution for low-volume, high-throughput sample processing that integrates liquid handling, DNA amplification, and real-time fluorescence detection in a single instrument. When combined with Douglas Scientific Array Tape™ and EnviroLogix DNAble®* chemistry, it provides a fast, high-quality alternative to real-time PCR for many research applications.

Chemistry

DNAble is a novel isothermal DNA amplification technique developed by EnviroLogix that utilizes two primers and a fluorescent molecular beacon to rapidly amplify and detect specific DNA sequences. Unlike PCR, DNAble uses two enzymes—a nicking enzyme and DNA polymerase—that work together at one temperature in the presence of a proprietary chemistry to amplify DNA specifically and exponentially. More importantly, real-time amplification curves can be generated and employed to quantify nucleic acid samples.

Amplifying a Human Pathogen

A dilution series spanning seven orders of magnitude was generated by diluting synthetic DNA that encodes the sequence of a common human pathogen. The dilution series used to build the standard curve was generated in just 10 minutes at 56°C. It utilized pathogen DNA in the DNAble assay with the Nexar system and a DNAble assay specific to that pathogen. The amplification curves are shown below in figure 1.

Figure 1. Amplification Curves

The regression line produced by the dilution series is shown in figure 2. Figure 2. Regression PlotThis figure clearly shows that the Nexar system is ideally suited to quantifying DNA, because the relationship between threshold time and quantity of DNA produced is virtually linear (r2 = 0.96).

More Information

To learn more about the Nexar Optimized for Real Time Nucleic Acid Processing and how it can lower your costs and significantly increase your sample throughput, go to www.douglasscientific.com.

*DNAble is a registered trademark of EnviroLogix, Inc.