Microfluidic nucleic acid analysis

摘要

Nucleic acid from cells and viruses sampled from a variety of environments may purified and expressed utilizing microfluidic techniques. In accordance with one embodiment of the present invention, individual or small groups of cells or viruses may be isolated in microfluidic chambers by dilution, sorting, and/or segmentation. The isolated cells or viruses may be lysed directly in the microfluidic chamber, and the resulting nucleic acid purified by exposure to affinity beads. Subsequent elution of the purified nucleic acid may be followed by ligation and cell transformation, all within the same microfluidic chip. In one specific application, cell isolation, lysis, and nucleic acid purification may be performed utilizing a highly parallelized microfluidic architecture to construct gDNA and cDNA libraries.

主权项

1. A method of obtaining nucleic acid from a cell sample, the method comprising:
(a) providing a sample comprising a plurality of cells suspended in solution; (b) physically isolating one or more cells of the sample within a microfluidic flow channel by a process that comprises flowing the solution partially through a first and a second valve in the microfluidic flow channel while the first and second valves are open, and then closing the first and second valves such that a part of the solution containing the one or more cells is isolated therebetween; (c) exposing the one or more cells in the part of the solution that was isolated between the first and the second valves in step (b) to a lysis chemical while said cell(s) are valved off from other cells in the sample, thereby releasing nucleic acid from the isolated cells; and (d) amplifying at least a portion of a nucleic acid released in step (c), wherein steps (b), (c) and (d) are performed in a microfluidic device.