Detection of hemi/homozygotes through heteroduplex formation in high-resolution melting analysis
| Autor: | Low Ps, Stacey K.H. Tay, Eric P.H. Yap, Jinting Cheng, Onn-Siong Yim, Poh San Lai |
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| Rok vydání: | 2011 |
| Předmět: |
Gene copy
Pcr cloning Gene Dosage Biophysics Single-nucleotide polymorphism Heteroduplex Analysis Biology Nucleic Acid Denaturation Polymerase Chain Reaction Polymorphism Single Nucleotide Biochemistry High Resolution Melt Humans Transition Temperature Copy-number variation Molecular Biology Heteroduplex formation Mixing (physics) Hemizygote Genetics Homozygote Nucleic Acid Heteroduplexes DNA Cell Biology Molecular biology Muscular Dystrophy Duchenne Dmd gene |
| Zdroj: | Analytical Biochemistry. 410:158-160 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2010.11.031 |
| Popis: | Heteroduplex formation, required for the complete detection of hemi/homozygotes using high-resolution melting analysis, can be induced either by pre-PCR mixing of genomic DNAs or by post-PCR mixing of PCR products from unknown and reference samples. This study investigates the effects of both methods using two single nucleotide polymorphisms in X-linked DMD gene. The results show that both methods resulted in the same effect when mixing samples with the same gene copy number. Mixing samples with different gene copy numbers has not been previously explored and we show that post-PCR mixing is insensitive to gene copy number differences as compared to pre-PCR mixing. |
| Databáze: | OpenAIRE |
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