{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T12:47:32Z","timestamp":1788871652363,"version":"build-2803163510"},"reference-count":15,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2007,8,1]],"date-time":"2007-08-01T00:00:00Z","timestamp":1185926400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Applied Biosystems"},{"name":"Corbett Life Science"},{"name":"Eppendorf"},{"name":"Idaho Technology"},{"DOI":"10.13039\/100015079","name":"ARUP Laboratories","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100015079","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2007,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Background: Additional instruments have become available since instruments for DNA melting analysis of PCR products for genotyping and mutation scanning were compared. We assessed the performance of these new instruments for genotyping and scanning for mutations.<\/jats:p>\n                  <jats:p>Methods: A 110-bp fragment of the \u03b2-globin gene including the sickle cell anemia locus (HBB c. 20A&amp;gt;T) was amplified by PCR in the presence of LCGreen Plus or SYBR Green I. Amplicons of 4 different genotypes [wild-type, homozygous, and heterozygous HBB c. 20A&amp;gt;T and double-heterozygote HBB c. (9C&amp;gt;T; 20A&amp;gt;T)] were melted on 7 different instruments [Applied Biosystems 7300, Corbett Life Sciences Rotor-Gene 6500HRM, Eppendorf Mastercycler RealPlex4S, Idaho Technology LightScanner (384 well), Roche LightCycler 480 (96 and 384 well) and Stratagene Mx3005p] at a rate of 0.61 \u00b0C\/s or when this was not possible, at 0.50 \u00b0C steps. We evaluated the ability of each instrument to genotype by melting temperature (Tm) and to scan for heterozygotes by curve shape.<\/jats:p>\n                  <jats:p>Results: The ability of most instruments to accurately genotype single-base changes by amplicon melting was limited by spatial temperature variation across the plate (SD of Tm = 0.020 to 0.264 \u00b0C). Other variables such as data density, signal-to-noise ratio, and melting rate also affected heterozygote scanning.<\/jats:p>\n                  <jats:p>Conclusions: Different instruments vary widely in their ability to genotype homozygous variants and scan for heterozygotes by whole amplicon melting analysis. Instruments specifically designed for high-resolution melting, however, displayed the least variation, suggesting better genotyping accuracy and scanning sensitivity and specificity.<\/jats:p>","DOI":"10.1373\/clinchem.2007.088120","type":"journal-article","created":{"date-parts":[[2007,6,7]],"date-time":"2007-06-07T21:11:13Z","timestamp":1181250673000},"page":"1544-1548","source":"Crossref","is-referenced-by-count":134,"title":["Expanded Instrument Comparison of Amplicon DNA Melting Analysis for Mutation Scanning and Genotyping"],"prefix":"10.1093","volume":"53","author":[{"given":"Mark G","family":"Herrmann","sequence":"first","affiliation":[{"name":"Institute for Clinical and Experimental Pathology, ARUP, Salt Lake City, UT;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob D","family":"Durtschi","sequence":"first","affiliation":[{"name":"Institute for Clinical and Experimental Pathology, ARUP, Salt Lake City, UT;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carl T","family":"Wittwer","sequence":"first","affiliation":[{"name":"Institute for Clinical and Experimental Pathology, ARUP, Salt Lake City, UT;"},{"name":"Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karl V","family":"Voelkerding","sequence":"first","affiliation":[{"name":"Institute for Clinical and Experimental Pathology, ARUP, Salt Lake City, UT;"},{"name":"Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2007,8,1]]},"reference":[{"key":"2020022802482197800_R1","doi-asserted-by":"crossref","unstructured":"Lay MJ, Wittwer CT. Real-time fluorescence genotyping of factor V Leiden during rapid-cycle PCR. Clin Chem1997;43:2262-2267.","DOI":"10.1093\/clinchem\/43.12.2262"},{"key":"2020022802482197800_R2","doi-asserted-by":"crossref","unstructured":"Krypuy M, Newnham GM, Thomas DM, Conron M, Dobrovic A. High resolution melting analysis for the rapid and sensitive detection of mutations in clinical samples: KRAS codon 12 and 13 mutations in non-small cell lung cancer. BMC Cancer2006;6:295.","DOI":"10.1186\/1471-2407-6-295"},{"key":"2020022802482197800_R3","doi-asserted-by":"crossref","unstructured":"Reed GH, Wittwer CT. Sensitivity and specificity of single-nucleotide polymorphism scanning by high-resolution melting analysis. Clin Chem2004;50:1748-1754.","DOI":"10.1373\/clinchem.2003.029751"},{"key":"2020022802482197800_R4","doi-asserted-by":"crossref","unstructured":"Zhou L, Vandersteen J, Wang L, Fuller T, Taylor M, Palais B, et al. High-resolution DNA melting curve analysis to establish HLA genotypic identity. Tissue Antigens2004;64:156-164.","DOI":"10.1111\/j.1399-0039.2004.00248.x"},{"key":"2020022802482197800_R5","doi-asserted-by":"crossref","unstructured":"Herrmann MG, Durtschi JD, Bromley LK, Wittwer CT, Voelkerding KV. Amplicon DNA melting analysis for mutation scanning and genotyping: cross-platform comparison of instruments and dyes. Clin Chem2006;52:494-503.","DOI":"10.1373\/clinchem.2005.063438"},{"key":"2020022802482197800_R6","doi-asserted-by":"crossref","unstructured":"Herrmann MG, Durtschi JD, Bromley LK, Wittwer CT, Voelkerding KV. Instrument comparison for heterozygote scanning of single and double heterozygotes: a correction and extension of Herrmann et al. Clin Chem2006;52:494-503Clin Chem 2007;53:150\u20132..","DOI":"10.1373\/clinchem.2006.081240"},{"key":"2020022802482197800_R7","doi-asserted-by":"crossref","unstructured":"Liew M, Pryor R, Palais R, Meadows C, Erali M, Lyon E, et al. Genotyping of single-nucleotide polymorphisms by high-resolution melting of small amplicons. Clin Chem2004;50:1156-1164.","DOI":"10.1373\/clinchem.2004.032136"},{"key":"2020022802482197800_R8","doi-asserted-by":"crossref","unstructured":"Graham R, Liew M, Meadows C, Lyon E, Wittwer CT. Distinguishing different DNA heterozygotes by high-resolution melting. Clin Chem2005;51:1295-1298.","DOI":"10.1373\/clinchem.2005.051516"},{"key":"2020022802482197800_R9","doi-asserted-by":"crossref","unstructured":"Gundry CN, Vandersteen JG, Reed GH, Pryor RJ, Chen J, Wittwer CT. Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotes. Clin Chem2003;49:396-406.","DOI":"10.1373\/49.3.396"},{"key":"2020022802482197800_R10","doi-asserted-by":"crossref","unstructured":"Palais RA, Liew MA, Wittwer CT. Quantitative heteroduplex analysis for single nucleotide polymorphism genotyping. Anal Biochem2005;346:167-175.","DOI":"10.1016\/j.ab.2005.08.010"},{"key":"2020022802482197800_R11","doi-asserted-by":"crossref","unstructured":"Wittwer CT, Reed GH, Gundry CN, Vandersteen JG, Pryor RJ. High-resolution genotyping by amplicon melting analysis using LCGreen. Clin Chem2003;49:853-860.","DOI":"10.1373\/49.6.853"},{"key":"2020022802482197800_R12","doi-asserted-by":"crossref","unstructured":"Zhou L, Wang L, Palais R, Pryor R, Wittwer CT. High-resolution DNA melting analysis for simultaneous mutation scanning and genotyping in solution. Clin Chem2005;51:1770-1777.","DOI":"10.1373\/clinchem.2005.054924"},{"key":"2020022802482197800_R13","doi-asserted-by":"crossref","unstructured":"Liew M, Seipp M, Durtschi J, Margraf RL, Dames S, Erali M, et al. Closed-tube SNP genotyping without labeled probes\/a comparison between unlabeled probe and amplicon melting. 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