Sodium bisulphite treatment of DNA combined with next generation sequencing (NGS) is a powerful combination for the interrogation of genome-wide DNA methylation profiles. Library preparation for whole genome bisulphite sequencing (WGBS) is challenging due to side effects of the bisulphite treatment, which leads to extensive DNA damage. Recently, a new generation of methods for bisulphite sequencing library preparation have been devised. They are based on initial bisulphite treatment of the DNA, followed by adaptor tagging of single stranded DNA fragments, and enable WGBS using low quantities of input DNA. In this study, we present a novel approach for quick and cost effective WGBS library preparation that is based on splinted adaptor tagging (SPLAT) of bisulphite-converted single-stranded DNA. Moreover, we validate SPLAT against three commercially available WGBS library preparation techniques, two of which are based on bisulphite treatment prior to adaptor tagging and one is a conventional WGBS method.
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http://dx.doi.org/10.1093/nar/gkw1110 | DOI Listing |
Food Chem
January 2025
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; State & Local Joint Engineering Research Center of Green Pesticide Invention and Application, Nanjing 210095, China. Electronic address:
Lateral flow immunoassay (LFIA) has the advantages of simplicity and rapidness, and is widely used for the rapid detection of pesticides and other analytes. However, small molecule compounds such as pesticides are often analyzed using competitive LFIA (CLFIA), whose sensitivity often does not meet the actual needs. In this study, a noncompetitive LFIA (NLFIA) for deltamethrin (DM) with high sensitivity was developed by using anti-immunocomplex peptides (AIcPs).
View Article and Find Full Text PDFAngew Chem Int Ed Engl
January 2025
Georgia State University, Chemistry, 50 Decatur ST SE, 30303, Atlanta, UNITED STATES OF AMERICA.
Poly-N-acetyllactosamine (poly-LacNAc) is ubiquitously expressed on cell surface glycoconjugates, serving as the backbone of complex glycans and an extended scaffold that presents diverse glycan epitopes. The branching of poly-LacNAc, where internal galactose (Gal) residues have β1-6 linked N-acetylglucosamine (GlcNAc) attached, forms the blood group I-antigen, which is closely associated with various physiological and pathological processes including cancer progression. However, the underlying mechanisms remain unclear as many of the I-antigen sequences are undefined and inaccessible.
View Article and Find Full Text PDFPopulations of very small embryonic-like stem cells (VSELs) (CD34+lin-CD45- and CD133+lin-CD45-), circulating in the peripheral blood of adults in small numbers, have been identified in several human tissues and together with the populations of hematopoietic stem cells (HSCs) (CD34+lin-CD45+) and CD133+lin-CD45+constitute a pool of cells with self-renewal and pluripotent stem cell characteristics. Using advanced cell staining and sorting strategies, we isolated populations of VSELs and HSCs for bulk RNA-Seq analysis to compare the transcriptomic profiles of both cell populations. Libraries were prepared from an extremely small number of cells; however, their good quality was preserved, and they met the criteria for sequencing.
View Article and Find Full Text PDFSTAR Protoc
January 2025
Division of Immunology and Infectious Disease Biology, INtegrative GENomics of HOst-PathogEn (INGEN-HOPE) Laboratory, CSIR-Institute of Genomics and Integrative Biology (CSIR-IGIB), Mall Road, Delhi 110007, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India. Electronic address:
Intracellular microorganisms like viruses and bacteria impact immune cell function. However, detection of these microbes is challenging as the majority exist in a non-culturable state. This protocol presents detailed steps to investigate intracellular microbial diversity using single-cell RNA sequencing (scRNA-seq) in immune-cells of SARS-CoV-2-positive and recovered patients.
View Article and Find Full Text PDFBackground: Liquid biopsy for Alzheimer's disease (AD) biomarkers in cerebral spinal fluid (CSF) or blood samples is becoming a significant sector in the management of AD and AD related dementia (ADRD). Whereas different types of molecules, such as small molecules, peptides, and antibodies, have been used as ligands for targeting AD biomarkers/pathologies such as amyloid beta peptides (Aβs), current liquid biopsy approaches mostly use antibodies. However, the activity and affinity of the antibodies can vary depending on the source and preparation as well as the stability in storage.
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