An efficient procedure for genotyping single nucleotide

Analysis of single nucleotide polymorphisms (SNPs) has been and will be increasingly utilized in various genetic disciplines, particularly in studying genetic determinants of complex diseases. Such studies will be facilitated by rapid, simple, low cost and high throughput methodologies for SNP genotyping. One such method is reported here, named tetra-primer ARMS-PCR, which employs two primer pairs to amplify, respectively, the two different alleles of a SNP in a single PCR reaction. A computer program for designing primers was developed. Tetra-primer ARMS-PCR was combined with microplate array diagonal gel electrophoresis, gaining the advantage of high throughput for gel-based resolution of tetraprimer ARMS-PCR products. The technique was applied to analyse a number of SNPs and the results were completely consistent with those from an independent method, restriction fragment length polymorphism analysis.

[1]  E. Thorland,et al.  Overlapping PCR for bidirectional PCR amplification of specific alleles: a rapid one-tube method for simultaneously differentiating homozygotes and heterozygotes. , 1997, Genome research.

[2]  R E Rhoads,et al.  Optimization of the annealing temperature for DNA amplification in vitro. , 1990, Nucleic acids research.

[3]  F. Collins,et al.  New goals for the U.S. Human Genome Project: 1998-2003. , 1998, Science.

[4]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[5]  T. Lehtimäki,et al.  Rapid identification of apolipoprotein E genotypes by multiplex amplification refractory mutation system PCR and capillary gel electrophoresis. , 1999, Clinical chemistry.

[6]  H. Blöcker,et al.  Predicting DNA duplex stability from the base sequence. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[7]  A. Brookes The essence of SNPs. , 1999, Gene.

[8]  R. Chiu,et al.  Determination of RhD zygosity: comparison of a double amplification refractory mutation system approach and a multiplex real-time quantitative PCR approach. , 2001, Clinical chemistry.

[9]  L. Brooks,et al.  A DNA polymorphism discovery resource for research on human genetic variation. , 1998, Genome research.

[10]  G. Lucotte,et al.  ARMS test for diagnosis of factor V Leiden mutation and allele frequencies in France. , 1998, Molecular and cellular probes.

[11]  D. Nickerson,et al.  Increasing the information content of STS-based genome maps: identifying polymorphisms in mapped STSs. , 1996, Genomics.

[12]  F. Green,et al.  Allele specific amplification by tetra-primer PCR. , 1992, Nucleic acids research.

[13]  C Summers,et al.  Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). , 1989, Nucleic acids research.

[14]  S. Humphries,et al.  Electrophoresis for genotyping: microtiter array diagonal gel electrophoresis on horizontal polyacrylamide gels, hydrolink, or agarose. , 1994, Analytical biochemistry.

[15]  B. Beghé,et al.  Comparison of three methods for single nucleotide polymorphism typing for DNA bank studies: Sequence‐specific oligonucleotide probe hybridisation, TaqMan liquid phase hybridisation, and microplate array diagonal gel electrophoresis (MADGE) , 1999, Human mutation.