US 8,133,670 B2Granted

Method for making populations of defined nucleic acid molecules

AssigneesCold Spring Harbor Laboratory, Merck & Co., Inc.

InventorsStephen H Friend, Michele A Cleary, Kristopher A Kilian, Ernest M Coffey, Gregory J Hannon, Patrick Paddison

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Abstract

The present invention provides methods of making a population of nucleic acid molecules, wherein each nucleic acid molecule comprises a predetermined nucleic acid sequence, each of said methods comprising the steps of: (a) synthesizing, on a substrate, a population of nucleic acid molecules wherein: i) each synthesized nucleic acid molecule comprises a predetermined nucleic acid sequence; and ii) each synthesized nucleic acid molecule is localized to a defined area of said substrate; (b) harvesting said population of synthesized nucleic acid molecules from said substrate to yield harvested nucleic acid molecules; and (c) introducing said harvested nucleic acid molecules into vector molecules.

Claims26

Dependent claims

A method for making a population of nucleic acid molecules, the method comprising:

(a) harvesting a population of synthesized single-stranded nucleic acid molecules that are covalently attached to a microarray, wherein each of the synthesized nucleic acid molecules consists of a predetermined nucleic acid sequence consisting of:
(i) a 5′ primer binding region,
(ii) a 3′ primer binding region,
(iii) one of a number of different target sequences located between the 5′ and 3′ primer binding regions, and additionally,
(iv) one of a number of unique target identifier sequences,
wherein the target identifier sequence is not comprised within the target sequence or primer binding regions, and
wherein each unique target identifier sequence is associated with only one particular target sequence;
(b) amplifying the population of synthesized single-stranded nucleic acid molecules harvested in step (a) to generate a population of double-stranded DNA molecules, and
(c) introducing the double-stranded DNA molecules into vector molecules.

6 more elements

Dependencies are inferred from the claim text.

Facts

Application: 10/860,559Filed 2 Jun 2004 · us provisional application
Priority: 13 Jun 2003
Published: US 2004/0259146 A123 Dec 2004
Granted: 13 Mar 2012
Term ends: Not estimatedThe source records don’t support an estimate.

Classification4 codes

C12Q 1/6837
Enzymatic or biochemical coupling of nucleic acids to a solid phase: using probe arrays or probe chips
Y10S 977/792
Molecular array: nucleic acid array, e.g. human genome array
C12Q
Measuring or testing processes involving enzymes, nucleic acids or microorganisms
Y10S
Technical subjects covered by former uspc cross-reference art collections [xracs] and digests