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SHR Neuro Cancer Cardio Lipid Metab Microb
Paar, C; Hammerl, V; Blessberger, H; Stekel, H; Steinwender, C; Berg, J.
Conditions of High Resolution Melting Analysis on the Cobas z480 Instrument for the Genotyping of VKORC1 in the Clinical Routine Laboratory.
Clin Lab. 2016; 62(12):2461-2467
Doi: 10.7754/Clin.Lab.2016.160616
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- Berg Jörg
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- Abstract:
- High resolution melting (HRM) of amplicons is a simple method for genotyping of single nucleotide polymorphisms (SNPs). Albeit many applications reported, HRM seems to be rarely used in clinical laboratories. The suitability of HRM-PCR for the clinical laboratory was investigated for genotyping of SNPs of the vitamin K epoxide reductase complex unit 1 gene. About 100 DNA samples were analyzed by two different HRM-PCRs on the Cobas z480 instrument and compared with a PCR with fluorescently labeled probes (HybProbe-PCR) on the LightCycler 2.0 instrument as reference. Reliable genotyping with 100% matching results was obtained, when the amplicon size was small (63 bp) and DNA input was limited by e.g., sample dilution with salt-free water. DNA extracted by differing methods may be used for genotyping by HRM-PCR. Compared with HybProbe-PCR, HRM-PCR on the Cobas z480 instrument allows for higher through-put, however, at the cost of a higher degree of laboratory standardization and a slower turnaround.
- Find related publications in this database (using NLM MeSH Indexing)
- Equipment Design -
- Genetic Testing - instrumentation
- Genetic Testing - methods
- Genotype -
- High-Throughput Nucleotide Sequencing - instrumentation
- Humans -
- Nucleic Acid Denaturation -
- Polymerase Chain Reaction - instrumentation
- Polymorphism, Single Nucleotide -
- Predictive Value of Tests -
- Reproducibility of Results -
- Vitamin K Epoxide Reductases - genetics
- Find related publications in this database (Keywords)
- genotyping
- SNP
- LightCycler
- real-time PCR
- FRET-probes
- high resolution melting
- HRM
- VKORC1