Low-temperature amplification
A new method of DNA amplification at 37°C
Scientists have developed a fast and easy-to–use method of DNA amplification at 37°C, which can greatly advance the field of DNA computing and molecular robotics, writes the Tokyo Institute of Technology with reference to Organic & Biomolecular Chemistry (Komiya et al., Leak-free million-fold DNA amplification with locked nucleic acid and targeted hybridization in one pot).
Scientists from the Tokyo Institute of Technology (Tokyo Tech) in Japan have developed a way to amplify DNA at a scale suitable for use in the emerging fields of DNA computing and molecular robotics. By providing highly sensitive detection of nucleic acids, their method can improve the diagnosis of diseases and accelerate the development of biosensors, for example, for food and the environment.
The method, called L-TEAM (low–temperature amplification), is the result of more than five years of research and offers a number of advantages over traditional PCR (polymerase chain reaction), the dominant technique used to amplify the DNA segments of interest.
Due to its ease of use, L-TEAM eliminates the need for heating and cooling stages and special equipment, which is usually required for PCR. Thus, it is an effective and inexpensive method that can significantly prevent protein denaturation, thereby opening the way to real-time analysis of living cells.
In their study, the scientists introduced synthetic molecules called blocked nucleic acids into DNA chains, as these molecules are known to help achieve greater stability during hybridization.
The addition of these nucleic acids led to an unexpected but useful result. The researchers observed a decrease in the level of "leakage" of amplification, which has long been a problem in DNA amplification studies, since this can lead to an error in the diagnosis of the disease, that is, to a false positive result.
"We were surprised to discover a new effect of blocked nucleic acids in overcoming the common problem of "leakage" in DNA amplification reactions," says Ken Komiya, associate professor at the Tokyo Technical School of Computing Technology. "We plan to investigate in detail the mechanisms of leak amplification and further improve the sensitivity and speed of the L–TEAM."
In the near future, this method may be used to detect short nucleic acids, such as microRNAs, for medical diagnostics. In particular, it could facilitate testing at the place of medical care and early detection of the disease. Nowadays, microRNAs are increasingly recognized as promising biomarkers for cancer detection and may hold the key to unlocking many other aspects of human health and environmental science.
In addition, Komiya explains that L-TEAM opens the way to the practical use of DNA computing and DNA-controlled molecular robotics. "The initial motivation for this work was to create a new enhanced module, which is necessary for building advanced molecular systems," he says. "Such systems can provide an understanding of the principle of action of living beings."
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