TY - JOUR
T1 - SBE-TAGS
T2 - An array-based method for efficient single-nucleotide polymorphism genotyping
AU - Hirschhorn, Joel N.
AU - Sklar, Pamela
AU - Lindblad-Toh, Kerstin
AU - Lim, Yin Mei
AU - Ruiz-Gutierrez, Melisa
AU - Bolk, Stacey
AU - Langhorst, Bradley
AU - Schaffner, Steven
AU - Winchester, Ellen
AU - Lander, Eric S.
PY - 2000/10/24
Y1 - 2000/10/24
N2 - Generating human single-nucleotide polymorphisms (SNPs) is no longer a rate-limiting step for genetic studies of disease. The number of SNPs in public databases already exceeds 200,000, and the total is expected to exceed 1,000,000 within a year. Rather, progress is limited by the inability to genotype large numbers of SNPs. Current genotyping methods are suitable for studying individual loci or at most a handful at a time. Here, we describe a method for parallel genotyping of SNPs, called single base extension-tag array on glass slides, SBE-TAGS. The principle is as follows. SNPs are genotyped by single base extension (SBE), using bifunctional primers carrying a unique sequence tag in addition to a locus-specific sequence. Because each locus has a distinct tag, the genotyping reactions can be performed in a highly multiplexed fashion, and the resulting product can then be 'demultiplexed' by hybridization to the reverse complements of the sequence tags arrayed on a glass slide. SBE-TAGS is simple and inexpensive because of the high degree of multiplexing and the use of an easily generated, generic tag array. The method is also highly accurate: we genotyped over 100 SNPs, obtaining over 5,000 genotypes, with approximately 99% accuracy.
AB - Generating human single-nucleotide polymorphisms (SNPs) is no longer a rate-limiting step for genetic studies of disease. The number of SNPs in public databases already exceeds 200,000, and the total is expected to exceed 1,000,000 within a year. Rather, progress is limited by the inability to genotype large numbers of SNPs. Current genotyping methods are suitable for studying individual loci or at most a handful at a time. Here, we describe a method for parallel genotyping of SNPs, called single base extension-tag array on glass slides, SBE-TAGS. The principle is as follows. SNPs are genotyped by single base extension (SBE), using bifunctional primers carrying a unique sequence tag in addition to a locus-specific sequence. Because each locus has a distinct tag, the genotyping reactions can be performed in a highly multiplexed fashion, and the resulting product can then be 'demultiplexed' by hybridization to the reverse complements of the sequence tags arrayed on a glass slide. SBE-TAGS is simple and inexpensive because of the high degree of multiplexing and the use of an easily generated, generic tag array. The method is also highly accurate: we genotyped over 100 SNPs, obtaining over 5,000 genotypes, with approximately 99% accuracy.
UR - http://www.scopus.com/inward/record.url?scp=12944305787&partnerID=8YFLogxK
U2 - 10.1073/pnas.210394597
DO - 10.1073/pnas.210394597
M3 - Article
C2 - 11035790
AN - SCOPUS:12944305787
SN - 0027-8424
VL - 97
SP - 12164
EP - 12169
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 22
ER -