Several specific problems are encountered when nonradioactive detection methods are used in conjunction with plant nucleic acids. In this chapter, we describe protocols for the isolation of DNA and RNA from plant leaves and the preparation of probe molecules by either PCR or in vitro transcription with different haptens. Furthermore, standard conditions and possible modifications for hybridization and detection of probes are given.
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http://dx.doi.org/10.1385/1-59745-229-7:69 | DOI Listing |
Curr Protoc
December 2024
Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.
Advances in sequencing technology have led to the discovery of diverse types of regulatory RNAs. Differential transcript levels regulate cellular processes and influence disease severity. Identifying these variations through reliable methods is crucial for understanding the regulatory roles and disease mechanisms of regulatory RNAs.
View Article and Find Full Text PDFJ Scleroderma Relat Disord
October 2024
Division of Diabetes, Endocrinology and Gastroenterology, Faculty of Biology, Medicine and Health, The University of Manchester, Northern Care Alliance NHS Foundation Trust, Salford Care Organisation, Manchester, UK.
Objective: Assessment of gastrointestinal and autonomic symptoms in patients with systemic sclerosis, and possible associations with gastric emptying rate.
Methods: Participant and patient disease-related characteristics were collected. Gastrointestinal and autonomic symptoms were assessed by the UCLA-SCTC GIT 2.
Methods Mol Biol
November 2024
Department of Microbiology and Molecular Medicine, Faculty of Medicine, University of Geneva, Institute of Genetics and Genomics of Geneva, Geneva, Switzerland.
Poly-A tail length dynamics have been extensively studied from yeast to human, mostly using reporter transcripts. Recent studies have been carried out genome-wide to determine the status of poly-A tails at steady state. However, poly-A tail measurement at equilibrium gives an overall length that reflects a mixture of the different poly-A tail sizes for a single transcript.
View Article and Find Full Text PDFMethods Mol Biol
November 2024
ncRNA, Epigenetic and Genome Fluidity, CNRS UMR3244, Sorbonne Université, PSL University, Institut Curie, Paris, France, Paris Cedex 05, France.
Northern blotting is a common technique in RNA biology, allowing to detect and quantify RNAs of interest following separation by gel electrophoresis, transfer to a membrane, and hybridization of specific anti-complementary labelled probes. In this chapter, we describe our protocol for efficient RNA extraction from yeast, separation on agarose gel, and capillary transfer to a membrane. We provide two different methods for strand-specific detection of several types of RNAs using oligonucleotide probes, the first using radioactive P-labelled probes, the second based on nonradioactive digoxigenin-labelled probes.
View Article and Find Full Text PDFPNAS Nexus
May 2023
Chemistry and Nanoscience Center, National Renewable Energy Laboratory, Golden, CO 80401, USA.
In hybrid perovskite solar cells (PSCs), the reaction of hydrogens (H) located in the amino group of the organic A-site cations with their neighboring halides plays a central role in degradation. Inspired by the retarded biological activities of cells in heavy water, we replaced the light H atom with its abundant, twice-as-heavy, nonradioactive isotope, deuterium (D) to hamper the motion of H. This D substitution retarded the formation kinetics of the detrimental H halides in Pb-based PSCs, as well as the H bond-mediated oxidation of Sn in Sn-Pb-based narrow-bandgap PSCs, evidenced by accelerated stability studies.
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