Unlabeled Oligonucleotides as Internal Temperature Controls for Genotyping by Amplicon Melting
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 کاملSNP genotyping by unlabeled probe melting analysis.
Fluorescent nucleic acid detection in polymerase chain reaction (PCR) generally uses oligonucleotide probes labeled with covalently attached dyes. However, unlabeled oligonucleotides in the presence of saturating DNA dyes can also serve as hybridization probes. The DNA dye, LCGreen Plus, and a 3'-blocked unlabeled probe are added before amplification, and asymmetric PCR is performed at a 1:5 to...
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...
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 کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: The Journal of Molecular Diagnostics
سال: 2007
ISSN: 1525-1578
DOI: 10.2353/jmoldx.2007.060136