A nonisotopic method for the detection of HIV DNA has been developed, based on the use of avidin-biotin complex to identify the products of polymerase chain reaction (PCR). Biotin label integrated in deoxyuridine triphosphate (bio-11-dUTP) was incorporated in the structure of amplified fragments in the course of PCR. The resultant products were identified in hybridization reaction with specific HIV DNA adsorbed on polystyrene plates. The suggested method was not inferior to the traditional radioisotope method in sensitivity and specificity and helped quantitatively assess the results of experiments using an objective criterion--optic density value.
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Mod Rheumatol
December 2024
Menzies Institute for Medical Research, University of Tasmania, Hobart, Australia.
Objectives: To describe the associations between OA-related biochemical markers and knee symptoms in middle-aged adults followed up over 10-13 years.
Methods: Blood samples were collected during the Childhood Determinants of Adult Health (CDAH)-1 study (year: 2004-06) and 10-13 year follow-up at CDAH-3. Serum samples from baseline (n=156) and follow-up (n=167) were analyzed for three OA-related biomarkers [cartilage oligomeric matrix protein (COMP), matrix metalloproteinase (MMP)-3, and hyaluronan (HA)] using non-isotopic enzyme-linked immunosorbent assay (ELISA).
J Vis Exp
June 2024
Department of Molecular Biology and Histocompatibility, General Hospital "Dr. Manuel Gea González";
The dot-blot is a simple, fast, sensitive, and versatile technique that enables the identification of minimal quantities of DNA specifically targeted by probe hybridization in the presence of carrier DNA. It is based on the transfer of a known amount of DNA onto an inert solid support, such as a nylon membrane, utilizing the dot-blot apparatus and without electrophoretic separation. Nylon membranes have the advantage of high nucleic acid binding capacity (400 µg/cm), high strength, and are positively or neutrally charged.
View Article and Find Full Text PDFInt J Mol Sci
February 2024
Laboratory of Poultry Medicine, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea.
Various chimeric lysins have been developed as efficacious antibiotics against multidrug-resistant bacteria, but direct comparisons of their antibacterial activities have been difficult due to the preparation of multiple recombinant chimeric lysins. Previously, we reported an cell-free expression method to better screen chimeric lysins against , but we still needed to increase the amounts of expressed proteins enough to be able to detect them non-isotopically for quantity comparisons. In this study, we improved the previous cell-free expression system by adding a previously reported artificial T7 terminator and reversing the different nucleotides between the T7 promoter and start codon to those of the T7 phage.
View Article and Find Full Text PDFEnviron Sci Pollut Res Int
March 2024
Unit Subsurface and Groundwater Systems, Deltares, Daltonlaan 600, Utrecht, 3484 BK, The Netherlands.
Understanding anaerobic biodegradation of ether oxygenates beyond MTBE in groundwater is important, given that it is replaced by ETBE as a gasoline additive in several regions. The lack of studies demonstrating anaerobic biodegradation of ETBE, and its product TBA, reflects the relative resistance of ethers and alcohols with a tertiary carbon atom to enzymatic attack under anoxic conditions. Anaerobic ETBE- or TBA-degrading microorganisms have not been characterized.
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