Single molecule quantitation and sequencing of rare translocations using microfluidic nested digital PCR.

ABSTRACT: Cancers are heterogeneous and genetically unstable. New methods are needed that provide the sensitivity and specificity to query single cells at the genetic loci that drive cancer progression, thereby enabling researchers to study the progression of individual tumors. Here, we report the development and application of a bead-based hemi-nested microfluidic droplet digital PCR (dPCR) technology to achieve 'quantitative' measurement and single-molecule sequencing of somatically acquired carcinogenic translocations at extremely low levels (<10(-6)) in healthy subjects. We use this technique in our healthy study population to determine the overall concentration of the t(14;18) translocation, which is strongly associated with follicular lymphoma. The nested dPCR approach improves the detection l

SUBMITTER: Shuga J 

PROVIDER: S-EPMC3763562 | biostudies-literature | 2013 Sep

REPOSITORIES: biostudies-literature