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US 7,521,186 B2Granted
PCR compatible nucleic acid sieving matrix
AssigneeCaliper Lifesciences Inc.
InventorTammy Burd Mehta
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Abstract
Sieving mediums comprising less than about 0.5% polymer, less than about 0.4% polymer, and 0.35% polymer or less are used to perform nucleic acid separations and PCR. The low polymer concentration does not inhibit PCR reactions and is sufficient for performing nucleic acids separations. Microfluidic devices are used to perform nucleic acids separations and PCR reactions in the sieving mediums described.
Claims9
A method of performing PCR and separating one or more PCR products, the method comprising:
- (i) mixing one or more PCR reaction components with an unpolymerized sieving medium in a channel of a microfluidic device to provide an unpolymerized PCR sieving medium within the channel;
- (ii) thermocycling the PCR sieving medium to produce one or more PCR products;
- (iii) polymerizing the sieving medium after thermocycling is completed, wherein the polymerized sieving medium has a polymer concentration that is less than 0.4%;
- and
- (iv) separating the one or more PCR products by flowing the one or more PCR products through the polymerized sieving medium.
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