We have developed a simple method for fast analysis of single nucleotide polymorphisms and identification of target clones from cloned complex PCR products. The method utilizes Kamchatka crab duplex-specific nuclease and universal fluorescent probe and is alternative to laborious screening procedures using radioactive probes, restriction analysis followed by gel electrophoresis or expensive sequencing. The method efficacy was demonstrated in several model experiments.
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http://dx.doi.org/10.1134/s1068162011040121 | DOI Listing |
Analyst
January 2025
School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei 230026, China.
MicroRNAs (miRNAs) are considered reliable biomarkers for a variety of diseases. However, their low abundance in organisms and high sequence similarity of homologous miRNAs make their accurate detection challenging. Here, we constructed a novel fluorescent biosensor for the detection of miRNA-155, a potential biomarker of neuroinflammation, based on duplex-specific nuclease (DSN) assisted amplification and DNA-templated silver nanoclusters (DNA-AgNCs) as fluorescence signal probes.
View Article and Find Full Text PDFZhongguo Zhong Yao Za Zhi
November 2024
Jilin Provincial Key Laboratory of Traditional Chinese Medicinal Materials Cultivation and Propagation, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences Changchun 130112, China College of Food and Biological Engineering, Chengdu University Chengdu 610106, China.
To construct a high-quality Panax ginseng cDNA library, transcription factors binding to the P. ginseng PgD14 gene promoter were screened by yeast one-hybrid, and proteins interacting with the P. ginseng Pgpht2-1 gene-encoded protein were screened by yeast two-hybrid.
View Article and Find Full Text PDFAnal Chem
December 2024
Department of Pharmacy, University of Naples "Federico II", 80131 Naples, Italy.
In the personalized medicine era, affordable and portable devices for quicker cancer monitoring, even in remote areas, are crucial. To address this need, we have developed an enzyme-assisted electrochemical point-of-care (POC) test for application toward liquid biopsy. In particular, miR-200a-5p has been taken into account as the model target due to its correlation for triple negative breast cancer (TNBC) prognosis.
View Article and Find Full Text PDFBiosensors (Basel)
November 2024
School of Materials Science and Engineering, Hainan University, Haikou 570228, China.
Fluorescent biosensor, which has the characteristics of high sensitivity, specificity, and low cost, can be directly detected in physiological fluids such as blood and serum. Therefore, the development of fluorescence sensor platforms for miRNA detection has a positive effect on the prevention and treatment of various diseases. In this paper, miR-34a was selected as a biological indicator of Alzheimer's disease (AD).
View Article and Find Full Text PDFTalanta
February 2025
School of Chemistry and Chemical Engineering, State Key Laboratory of Digital Medical Engineering, Southeast University, Nanjing, 211189, China. Electronic address:
Alkaline phosphatase (ALP) is an essential hydrolase widely present in humans, and it extensively acts as a biomarker for multiple human diseases. Conventional ALP assays suffer from complicated synthesis, tedious operation, low sensitivity, and large sample consumption. Herein, we construct an end-repairing-engineered quadratic in vitro transcription machine for single-molecule monitoring of ALP in diverse cancers with 3'-phosphoryl (PO) nucleic acid as a macromolecular substrate.
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