Genotypic analysis of multiple loci in somatic cells by whole genome amplification.
To screen multiple loci in small purified samples of diploid and aneuploid cells a PCR-based technique of whole genome amplification was adapted to the study of somatic lesions. DNA samples from different numbers of flow-sorted diploid and aneuploid cells from biopsies were amplified with a degenerate 15mer primer. Aliquots of these reactions were then used in locus-specific reactions using a single round of PCR cycles with individual sets of primers representing polymorphic markers for different regions. As a result, polymorphic markers for different chromosomal regions, including VNTRs and dinucleotide repeats, can be used to perform up to 30 locus-specific PCR assays with a single sample obtained from fewer than 1000 cells.
Barrett
M T
MT
Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USA.
Reid
B J
BJ
Joslyn
G
G
D000230:Adenocarcinoma / Q000235:genetics*; D000483:Alleles; D004273:DNA, Neoplasm / Q000235:genetics*; D004938:Esophageal Neoplasms / Q000235:genetics*; D005819:Genetic Markers; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D017384:Sequence Deletion
D004273:DNA, Neoplasm; D005819:Genetic Markers