{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T12:40:08Z","timestamp":1740141608761,"version":"3.37.3"},"reference-count":26,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2005,2,23]],"date-time":"2005-02-23T00:00:00Z","timestamp":1109116800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Danish Sue Program","award":["9800647-67"],"award-info":[{"award-number":["9800647-67"]}]},{"DOI":"10.13039\/100008392","name":"Danish Medical Research Council","doi-asserted-by":"crossref","award":["9600259"],"award-info":[{"award-number":["9600259"]}],"id":[{"id":"10.13039\/100008392","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Harboe Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2005,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Single nucleotide polymorphisms (SNPs) represent the most frequent form of genetic variations. Some of the most sensitive methods for SNP genotyping employ synthetic oligonucleotides, such as the peptide nucleic acid (PNA). We introduce a new method combining allele\u2010specific hybridization, PNA technology, and flow cytometric detection. We tested the design by genotyping a Danish basal cell carcinoma cohort of 80 individuals for an <jats:italic>A<\/jats:italic>\/<jats:italic>C<\/jats:italic> SNP in exon 6 of the <jats:italic>XPD<\/jats:italic> gene.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Genomic DNA was amplified by a two\u2010step polymerase chain reaction (PCR) in the presence of fluorescein\u2010dyed primers and fluorescein\u201012\u2010dUTP. The allele\u2010specific PNA molecules were covalently coupled to carboxylated microspheres with and without rhodamine. Allele\u2010specific hybridization between PCR products and immobilized PNA was carried out at 60\u00b0C followed by flow cytometric detection.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We present a fully functional two\u2010bead genotyping system based on PNA capture and flow cytometric detection used for the correct and fast regenotyping of a Danish basal cell carcinoma cohort.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This new assay presents a simple, rapid, and robust method for SNP genotyping for laboratories equipped with a standard flow cytometer. Moreover, this system offers potential for multiplexing and will be operational for middle\u2010scale genotyping. \u00a9 2005 Wiley\u2010Liss, Inc.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.a.20123","type":"journal-article","created":{"date-parts":[[2005,2,23]],"date-time":"2005-02-23T19:41:27Z","timestamp":1109187687000},"page":"80-86","source":"Crossref","is-referenced-by-count":14,"title":["SNP genotyping using microsphere\u2010linked PNA and flow cytometric detection"],"prefix":"10.1002","volume":"64A","author":[{"given":"Eszter","family":"Rockenbauer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenneth","family":"Petersen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ulla","family":"Vogel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lars","family":"Bolund","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steen","family":"K\u00f8lvraa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kirsten Vang","family":"Nielsen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bj\u00f8rn A.","family":"Nex\u00f8","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,2,23]]},"reference":[{"key":"e_1_2_6_2_2","first-page":"43","article-title":"Detection of single nucleotide polymorphisms","volume":"5","author":"Kwok PY","year":"2003","journal-title":"Curr Issues Mol 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