Highly sensitive and multiplexed one-step RT-qPCR for profiling genes involved in the circadian rhythm using microparticles
- Authors
- Kim, M.Y.; Jung, S.; Kim, J.; Lee, H.J.; Jeong, S.; Sim, S.J.; Kim, S.K.
- Issue Date
- 19-3월-2021
- Publisher
- Nature Research
- Citation
- Scientific Reports, v.11, no.1
- Indexed
- SCIE
SCOPUS
- Journal Title
- Scientific Reports
- Volume
- 11
- Number
- 1
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/129053
- DOI
- 10.1038/s41598-021-85728-y
- ISSN
- 2045-2322
- Abstract
- Given the growing interest in molecular diagnosis, highly extensive and selective detection of genetic targets from a very limited amount of samples is in high demand. We demonstrated the highly sensitive and multiplexed one-step RT-qPCR platform for RNA analysis using microparticles as individual reactors. Those particles are equipped with a controlled release system of thermo-responsive materials, and are able to capture RNA targets inside. The particle-based assay can successfully quantify multiple target RNAs from only 200 pg of total RNA. The assay can also quantify target RNAs from a single cell with the aid of a pre-concentration process. We carried out 8-plex one-step RT-qPCR using tens of microparticles, which allowed extensive mRNA profiling. The circadian cycles were shown by the multiplex one-step RT-qPCR in human cell and human hair follicles. Reliable 24-plex one-step RT-qPCR was developed using a single operation in a PCR chip without any loss of performance (i.e., selectivity and sensitivity), even from a single hair. Many other disease-related transcripts can be monitored using this versatile platform. It can also be used non–invasively for samples obtained in clinics. © 2021, The Author(s).
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Collections - Graduate School > Department of Biomedical Sciences > 1. Journal Articles
- College of Engineering > Department of Chemical and Biological Engineering > 1. Journal Articles
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