Megapixel digital PCR

We present a microfluidic 'megapixel' digital PCR device that uses surface tension–based sample partitioning and dehydration control to enable high-fidelity single DNA molecule amplification in 1,000,000 reactors of picoliter volume with densities up to 440,000 reactors cm−2. This device achieves a dynamic range of 107, single-nucleotide-variant detection below one copy per 100,000 wild-type sequences and the discrimination of a 1% difference in chromosome copy number.

[1]  F. Young.  Biochemistry , 1955, The Indian Medical Gazette.

[2]  T. Kunkel,et al.  Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. , 1988, Biochemistry.

[3]  G. Whitesides,et al.  Fabricating large arrays of microwells with arbitrary dimensions and filling them using discontinuous dewetting. , 1998, Analytical chemistry.

[4]  K. Kinzler,et al.  Digital PCR. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[6]  S. Quake,et al.  A robust and scalable microfluidic metering method that allows protein crystal growth by free interface diffusion , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Frank Diehl,et al.  BEAMing: single-molecule PCR on microparticles in water-in-oil emulsions , 2006, Nature Methods.

[8]  David Bryder,et al.  Transcription factor profiling in individual hematopoietic progenitors by digital RT-PCR , 2006, Proceedings of the National Academy of Sciences.

[9]  Frank Diehl,et al.  BEAMing up for detection and quantification of rare sequence variants , 2006, Nature Methods.

[10]  P. Brousset,et al.  Frequent detection of the JAK2 V617F mutation in bone marrow core biopsy specimens from chronic myeloproliferative disorders using the TaqMan polymerase chain reaction single nucleotide polymorphism genotyping assay: a retrospective study with pathologic correlations. , 2006, Human pathology.

[11]  Bill W Colston,et al.  High-throughput quantitative polymerase chain reaction in picoliter droplets. , 2008, Analytical chemistry.

[12]  S. Goodman,et al.  Circulating mutant DNA to assess tumor dynamics , 2008, Nature Medicine.

[13]  H. C. Fan,et al.  Noninvasive diagnosis of fetal aneuploidy by shotgun sequencing DNA from maternal blood , 2008, Proceedings of the National Academy of Sciences.

[14]  Rustem F Ismagilov,et al.  Digital PCR on a SlipChip. , 2010, Lab on a chip.

[15]  Bruce K Gale,et al.  Spinning disk platform for microfluidic digital polymerase chain reaction. , 2010, Analytical chemistry.