用于罕见序列变异检测和定量的BEAMing技术

BEAMing up for detection and quantification of rare sequence variants.

作者信息

Li Meng, Diehl Frank, Dressman Devin, Vogelstein Bert, Kinzler Kenneth W

机构信息

The Sidney Kimmel Comprehensive Cancer Center, Howard Hughes Medical Institute, and Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, 1650 Orleans Street, Baltimore, Maryland 21231, USA.

出版信息

Nat Methods. 2006 Feb;3(2):95-7. doi: 10.1038/nmeth850.

DOI:10.1038/nmeth850

PMID:16432518

Abstract

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.

摘要

BEAMing技术可实现将一群DNA片段一对一转化为一群磁珠。我们利用滚环扩增技术，使结合在这些磁珠上的拷贝数增加了100多倍。即使突变型和野生型序列的比例低于1:10,000，也能对其进行计数，并且灵敏度足以直接定量用于PCR的DNA聚合酶的错误率。