High-Speed Melting Analysis: The Effect of Melting Rate on Small Amplicon Microfluidic Genotyping
نویسندگان
چکیده
منابع مشابه
Multiplex amplicon genotyping by high-resolution melting.
High-resolution amplicon melting is a simple method for genotyping that uses only generic PCR primers and a saturating DNA dye. Multiplex amplicon genotyping has previously been reported in a single color, but two instruments were required: a carousel-based rapid cycler and a high-resolution melting instrument for capillaries. Manual transfer of capillaries between instruments and sequential me...
متن کاملHigh-resolution genotyping by amplicon melting analysis using LCGreen.
BACKGROUND High-resolution amplicon melting analysis was recently introduced as a closed-tube method for genotyping and mutation scanning (Gundry et al. Clin Chem 2003;49:396-406). The technique required a fluorescently labeled primer and was limited to the detection of mutations residing in the melting domain of the labeled primer. Our aim was to develop a closed-tube system for genotyping and...
متن کاملMicrofluidic genotyping by rapid serial PCR and high-speed melting analysis.
BACKGROUND Clinical molecular testing typically batches samples to minimize costs or uses multiplex lab-on-a-chip disposables to analyze a few targets. In genetics, multiple variants need to be analyzed, and different work flows that rapidly analyze multiple loci in a few targets are attractive. METHODS We used a microfluidic platform tailored to rapid serial PCR and high-speed melting (HSM) ...
متن کاملLimitations of genotyping based on amplicon melting temperature.
interferences”. We considered the latter two sources of error to be outside the scope of our study, in part because it is difficult to know how one might model the interferences. We did, however, discuss the importance of the continuing efforts of manufacturers to design instruments that avoid sources of error, such as those encountered by patients with special needs. We believe that our conclu...
متن کاملSmall amplicon genotyping using internal temperature calibration and high resolution-melting.
Small amplicon genotyping is a simple alternative to probe-based methods and more rapid and cost-effective than sequencing. This approach relies on the thermodynamic differences between two molecules that differ by potentially only a single base. Because there are no custom probes (labeled or unlabeled) required, assay design and optimization are simplified, making this technique rapid and inex...
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ژورنال
عنوان ژورنال: Clinical Chemistry
سال: 2017
ISSN: 0009-9147,1530-8561
DOI: 10.1373/clinchem.2017.276147