Detection of hemi/homozygotes through heteroduplex formation in high-resolution melting analysis.
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.
Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119074, Singapore.
Jinting
J
Cheng
Onn-Siong
OS
Yim
Poh-Sim
PS
Low
Stacey K H
SK
Tay
Eric P H
EP
Yap
Poh-San
PS
Lai
D004247:DNA / Q000737:chemistry / Q000235:genetics; D018628:Gene Dosage; D057898:Hemizygote; D020180:Heteroduplex Analysis; D006720:Homozygote; D006801:Humans; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D044366:Transition Temperature
D009692:Nucleic Acid Heteroduplexes; D004247:DNA