US 7,482,443 B2
	Systems and methods to quantify and amplify both signaling probes for cDNA chips and genes expression microarrays
David A. Shafer, Atlanta, Ga. (US)
Assigned to Genetag Technology, Inc.,
Appl. No. 10/380,596 PCT Filed Mar. 09, 2001, PCT No. PCT/US01/07508 § 371(c)(1), (2), (4) Date Mar. 17, 2003, PCT Pub. No. WO01/66802, PCT Pub. Date Sep. 13, 2001.
Application 10/380596 is a continuation in part of application No. 09/744097, filed on Jan. 16, 2001.
Claims priority of provisional application 60/187982, filed on Mar. 09, 2000.
Prior Publication US 2004/0053275 A1, Mar. 18, 2004
Int. Cl. C07H 21/04 (2006.01); C12Q 1/68 (2006.01); C12P 19/34 (2006.01)

U.S. Cl. 536—24.3  [435/6; 435/91.1; 435/91.2]

10 Claims

1. A probe set composition for the detection of genomic sequences, to provide terminal signaling per probe and to enable global amplification, global labeling and global addition of reporters to said probe set, wherein said probe set composition comprises a pool of modified cDNA probes, wherein each probe comprises, as a central target-specific sequence, a single truncated cDNA segment comprising a copy of the 3′ end of each mRNA transcript or an amplified set of said probes, wherein each probe further comprises an appended terminal tri-functional sequence on one or both ends; wherein said sequence comprises at least one of SEQ ID NO: 1 and its complement, SEQ ID NO: 2 and its complement, SEQ ID NO: 3 and its complement or SEQ ID NO: 4 and its complement, and wherein the tri-functional sequence is: (i) a common universal linker for binding a common reporter unit to each probe of the probe set, wherein each reporter unit comprises at least one terminal single stranded polynucleotide linker complementary to the universal linker of the members of the probe set and an attached reporter segment, preferentially comprising double stranded DNA, that additionally comprises at least two labeling molecules, or (ii) a common universal primer binding site for globally copying and amplifying the probe set, or (iii) a common universal reporter binding site wherein the universal reporter comprises a single-stranded polynucleotide that is complementary to the universal reporter binding site and that additionally comprises at least one labeling compound that is directly attached to the 5′ end or to an internal base of said polynucleotide, and wherein said labeled polynucleotides are incorporated into the members of the probe set during amplification and/or appended to the members of the probe set by hybridization.