The polymerase chain reaction (PCR) can be used to produce both nonradiolabeled DNA probes and radiolabeled DNA probes with high specific activity. In this protocol, PCR is used to generate double-stranded probes. Related methods, including the generation of asymmetric probes by PCR, are also discussed.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1101/pdb.prot100610 | DOI Listing |
Hortic Res
January 2025
Key Laboratory of Xinjiang Phytomedicine Resource and Utilization of Ministry of Education, Key Laboratory of Oasis Town and Mountain-basin System Ecology of Xinjiang Production and Construction Corps, College of Life Sciences, Shihezi University, Shihezi 832003, China.
Alfalfa is one of the most economically valuable forage crops in the world. However, molecular cytogenetic studies in alfalfa lag far behind other cash crops and have reached a bottleneck. Here, we developed a novel chromosome identification system by designing 21 oligo probes in specific regions of each chromosome, which can be used as a barcode to simultaneously distinguish all chromosomes in a cell.
View Article and Find Full Text PDFJ Inflamm Res
January 2025
Key Laboratory of Biomechanics and Mechanobiology, Ministry of Education; Key Laboratory of Innovation and Transformation of Advanced Medical Devices, Ministry of Industry and Information Technology; National Medical Innovation Platform for Industry-Education Integration in Advanced Medical Devices (Interdiscipline of Medicine and Engineering); School of Engineering Medicine, Beihang University, Beijing, 100191, People's Republic of China.
Background: Lichen sclerosus (LS) is a chronic inflammatory disease affecting skin and mucosal tissues, particularly external genitalia, with a risk of cancer. Its etiology is unknown, possibly involving immune dysregulation and inflammation.
Methods: Study used DNA methylation (DNAme) and single-cell RNA sequencing (scRNA-seq) to compare LS with normal skin.
Anal Chem
January 2025
Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Aptamer Engineering Center of Hunan Province, Hunan University, Changsha, Hunan 410082, China.
DNA reaction equilibrium-based calculations have great potential in thermodynamic characterization, but their widespread applications are hindered by significant measurement deviation of equilibrium concentration. Here, we report the advantages of metastable DNA hybridization in reducing quantification deviation of equilibrium concentration and propose a universal and standardized strategy for measuring aptamer binding energy, termed metastable DNA reference calorimetry (MDRC). We built different MDRC-based algorithms tailored to different aptamer binding models, enabling the calculation of thermodynamic parameters for aptamers with one or more binding sites.
View Article and Find Full Text PDFAnal Chim Acta
February 2025
School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, China. Electronic address:
Background: DNA methylation catalyzed by various DNA methyltransferases (DNA MTases) is one of the important epigenetic regulations in both eukaryotes and prokaryotes. Therefore, the detection of DNA MTase activity is a vital target and direction in the study of methylation-related diseases.
Results: In this study, an ultrasensitive and robust strategy was developed for DNA MTase activity sensing based on bifunctional probe propelling multipath strand displacement amplification and CRISPR/Cas12a techniques.
Talanta
January 2025
Department of Chemical Sciences, University of Catania, Viale Andrea Doria 6, 95122, Catania, Italy; INBB, Istituto Nazionale di Biostrutture e Biosistemi, Viale delle Medaglie d'Oro, 305, 00136, Roma, Italy. Electronic address:
Directly detecting biomarkers in liquid biopsy for diagnosis and personalized treatment plays a crucial role in managing cancer relapse and increasing survival rates. Typically, the standard analysis of circulating tumour DNA requires lengthy isolation, extraction, and amplification steps, leading to sample contamination, longer turnaround time and higher assay costs. Surface plasmon resonance is an emerging and promising technology for rapid and real-time dynamic biomarker monitoring in liquid biopsy.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!