BEAMing up for detection and quantification of rare sequence variants

• Published in: Nature methods Vol. 3; no. 2; pp. 95 - 97
• Main Authors: Vogelstein, Bert, Kinzler, Kenneth W, Li, Meng, Diehl, Frank, Dressman, Devin
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01-02-2006
• Subjects: Deoxyribonucleic acid; DNA; DNA - analysis; DNA - genetics; DNA - metabolism; DNA Mutational Analysis - methods; DNA polymerases; DNA-Directed DNA Polymerase - metabolism; Emulsions - chemistry; Exons - genetics; Flow Cytometry; Fluorescent Dyes - chemistry; Genetic variation; Identification and classification; Measurement; Microscopy, Fluorescence; Microspheres; Mutation - genetics; Nucleic Acid Amplification Techniques - methods; Oils - chemistry; Physiological aspects; Polymerase chain reaction; Polymerase Chain Reaction - methods; Reproducibility of Results; Tumor Suppressor Protein p53 - genetics; Water - chemistry; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth850

BEAMing allows the one-to-one conversion of a population of DNA fragments into a population of beads. We used rolling circle amplification to increase the number of copies bound to such beads by more than 100-fold. This allowed enumeration of mutant and wild-type sequences even when they were present at ratios less than 1:10,000 and was sensitive enough to directly quantify the error rate of DNA polymerases used for PCR.