Method of sampling, amplifying and quantifying segment of nucleic acid, polymerase chain reaction assembly having nanoliter-sized sample chambers, and method of filling assembly
First Claim
1. An assembly for containing a portion of fluid material, said assembly comprising:
- a substrate and a cover having facing surfaces spaced from one another, the facing surface of at least one of said substrate and cover comprising a first material;
a flow-through channel between said substrate and said cover; and
a first fluid retaining means located in said flow-through channel and being in communication with said flow-through channel, said first fluid retaining means having a first affinity to a first fluid and a second affinity to a displacing fluid, said flow through channel having a third affinity to said first fluid and a fourth affinity to said displacing fluid, wherein said first, second, third, and fourth affinities are such that;
(1) the first fluid retaining means retains at least a portion of the first fluid when the first fluid is loaded into the assembly; and
(2) the displacing fluid displaces first fluid from the flow-through channel without substantially displacing first fluid from the first fluid retaining means when the displacing fluid is loaded into the assembly after the first fluid is loaded into the assembly.
5 Assignments
0 Petitions
Accused Products
Abstract
Methods of detecting and quantifying genomic nucliec acid molecule sequences are provided using the simultaneous amplification of a plurality of discrete nanoliter-sized samples. A miniaturized closed assembly is also provided for carrying out amplification of a nucleic acid molecule by polymerase chain reaction in multiple nanoliter-sized samples. Methods of filling miniaturized sample chambers are also provided as are methods for determining the number of template molecules in a sample by conducting replicate nucleic acid sequence amplification reactions on a set of terminally diluted samples and counting the number of positive amplification reactions. The methods can be used to detect a single starting nucleic acid target molecule.
-
Citations
22 Claims
-
1. An assembly for containing a portion of fluid material, said assembly comprising:
- a substrate and a cover having facing surfaces spaced from one another, the facing surface of at least one of said substrate and cover comprising a first material;
a flow-through channel between said substrate and said cover; and
a first fluid retaining means located in said flow-through channel and being in communication with said flow-through channel, said first fluid retaining means having a first affinity to a first fluid and a second affinity to a displacing fluid, said flow through channel having a third affinity to said first fluid and a fourth affinity to said displacing fluid, wherein said first, second, third, and fourth affinities are such that;
(1) the first fluid retaining means retains at least a portion of the first fluid when the first fluid is loaded into the assembly; and
(2) the displacing fluid displaces first fluid from the flow-through channel without substantially displacing first fluid from the first fluid retaining means when the displacing fluid is loaded into the assembly after the first fluid is loaded into the assembly. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
- a substrate and a cover having facing surfaces spaced from one another, the facing surface of at least one of said substrate and cover comprising a first material;
Specification