Direct molecular haplotyping of long-range genomic DNA with M1-PCR.
Haplotypes, combinations of several phase-determined polymorphic markers, are extremely valuable for studies of disease association and chromosome evolution. Here we describe a technique called M1-PCR (M for "multiplex" and 1 for "single-copy DNA molecules") that enables direct molecular haplotyping of several polymorphic markers separated by as many as 24 kb. A genomic DNA sample first is diluted to approximately single-copy. The haplotype is directly determined by simultaneously genotyping several polymorphic markers in the same reaction with a multiplex PCR and base extension reaction. This approach does not rely on pedigree data and does not require previous amplification of the entire genomic region containing the selected markers.
Ding
Chunming
C
Bioinformatics Program and Center for Advanced Biotechnology, Boston University, 36 Cummington Street, Boston, MA 02215, USA.
Cantor
Charles R
CR
D002352:Carrier Proteins / Q000235:genetics; D053480:Cholesterol Ester Transfer Proteins; D004247:DNA / Q000032:analysis*; D017931:DNA Primers / Q000737:chemistry; D005821:Genetic Techniques; D015894:Genome, Human; D005838:Genotype; D006023:Glycoproteins; D006239:Haplotypes; D006801:Humans; D008957:Models, Genetic; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*
C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D017931:DNA Primers; D006023:Glycoproteins; D004247:DNA