Genotyping accuracy of high-resolution DNA melting instruments

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Publication Type pre-print
School or College School of Medicine
Department Pathology
Creator Wittwer, Carl T.
Other Author Li, Mei.; Zhou, Luming; Palais, Robert A.
Title Genotyping accuracy of high-resolution DNA melting instruments
Date 2014-01-01
Description High-resolution DNA melting is a closed-tube method for genotyping and variant scanning that depends on the thermal stability of PCR-generated products. Instruments vary in thermal precision, sample format, melting rates, acquisition, and software. Instrument genotyping accuracy has not been assessed. Each genotype of the single nucleotide variant (SNV) (c.3405-29A>T) of CPS1 (carbamoyl-phosphate synthase 1, mitochondrial) was amplified by PCR in the presence of LCGreen Plus with 4 PCR product lengths. After blinding and genotype randomization, samples were melted in 10 instrument configurations under conditions recommended by the manufacturer. For each configuration and PCR product length, we analyzed 32-96 samples (depending on batch size) with both commercial and custom software. We assessed the accuracy of heterozygote detection and homozygote differentiation of a difficult, nearest- neighbor symmetric, class 4 variant with predicted <Tmof 0.00 °C.
Type Text
Publisher American Association for Clinical Chemistry
Volume 60
Issue 6
First Page 864
Last Page 872
Language eng
Bibliographic Citation Li, M., Zhou, L., Palais, R. A., & Wittwer, C. T. (2014). Genotyping accuracy of high-resolution DNA melting instruments. Clinical Chemistry, 60(6), 864-72.
Rights Management (c)American Association for Clinical Chemistry
Format Medium application/pdf
Format Extent 1,308,283 bytes
Identifier uspace,18737
ARK ark:/87278/s6cc48w6
Setname ir_uspace
Date Created 2016-06-10
Date Modified 2021-05-06
ID 713374
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