Single-tube genotyping from a human hair root by direct PCR.
We have developed a simple, labor-saving, inexpensive and rapid SNP genotyping method that directly uses a human hair root as the template. This single-tube genotyping method was used to successfully and reliably genotype the ADH1B and ALDH2 polymorphisms using a hair root (without DNA isolation) and the polymerase chain reaction (PCR) enzyme kit KOD FX. Since the DNA extraction step was eliminated, the possibility of sample contamination was considerably decreased. The single-tube SNP genotyping was performed by coupling the PCR enzyme kit with allele-specific primer (ASP)-PCR. In the post-genome era, a simple and inexpensive method for diagnostic analysis is in high demand, and this PCR method with a hair root as a template will be very useful for genetic diagnoses in biological and medical laboratories.
Hayashida
Mariko
M
School of Pharmaceutical Sciences, Mukogawa Women's University, Koshien, Nishinomiya 663-8179, Japan.
Iwao-Koizumi
Kyoko
K
Murata
Shigenori
S
Kinoshita
Kenji
K
D000293:Adolescent; D000426:Alcohol Dehydrogenase / Q000235:genetics; D000444:Aldehyde Dehydrogenase / Q000235:genetics; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D005260:Female; D005838:Genotype; D006197:Hair / Q000033:anatomy & histology* / Q000378:metabolism*; D006801:Humans; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D013997:Time Factors; D055815:Young Adult
C489332:ADH1B protein, human; D000426:Alcohol Dehydrogenase; C096127:ALDH2 protein, human; D000444:Aldehyde Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial