By Nate Chongsiriwatana --
Previously, single-molecule DNA analysis has been limited by the difficulty of linearizing (untangling) and manipulating intact native DNA. To address these limitations, a Princeton University research team, including Dr. Han Cao, the founder and chief scientific officer of BioNanomatrix, used a nanofluidic chip to untangle and guide individual DNA molecules into an array of nanochannels. According to Dr. Cao, the founder and chief scientific officer of BioNanomatrix, "this invention allows for true linear analysis of very long biomolecules such as native genomic DNA hundreds of thousands of base pairs in length without cloning or PCR amplification." This technique, which is the basis for the company's NanoAnalyzer® system, can be used for multiplexed parallel processing applications, such as direct imaging analysis of structural variations in a person's genome and DNA mapping and sequencing.
At the crux of the invention are three critical dimensional qualities for nanochannel arrays that enable the simultaneous isolation and analysis of a multitude of elongated macromolecules. Specifically, the '770 patent discloses that: (1) the channels should be sufficiently narrow in order to elongate and isolate macromolecules; (2) the channels should be long enough to permit the instantaneous observation of the entire elongated macromolecule; and (3) a high number of channels should be provided in each array to permit the simultaneous observation of a large number of macromolecules.
The lone independent claim of the '770 patent
1. A method of isolating, imaging, and analyzing, in parallel, two or more individual nucleic acid biopolymers, comprising the steps of: providing a nanofluidic chip, comprising: a) nanochannel array, comprising: a surface having a plurality of parallel nanochannels running along the material of the surface, said nanochannels having a trench width of less than about 150 nanometers and a trench depth of less than 200 nanometers; at least one of the plurality of nanochannels being surmounted by sealing material to render such nanochannels at least substantially enclosed; at least two of the nanochannels capable of admitting a fluid; b) at least one sample reservoir in fluid communication with at least two of the nanochannels, said sample reservoir capable of releasing a fluid containing at least two nucleic acid biopolymer; providing the at least one sample reservoir with at least one fluid, said fluid comprising at least two nucleic acid biopolymers; transporting the at least two nucleic acid biopolymers into the at least two nanochannels to elongate said at least two nucleic acid biopolymers, the at least two nucleic acid biopolymers being individually confined within the at least two nanochannels such that the at least two nucleic acid biopolymer are transported through the at least two nanochannels in an unfolded fashion; imaging in parallel at least one signal transmitted from the at least two elongated and isolated nucleic acid biopolymers within the nanochannels; and correlating the signal to at least one property of the at least two nucleic acid biopolymers to thereby analyze the two or more individual nucleic acid biopolymers.
The specification also discloses methods of preparing nanochannel arrays and nanofluidic chips, methods of analyzing macromolecules, such as entire strands of genomic DNA, and systems for carrying out these methods.