Method for serial analysis of gene expression

• US 20030049653A1
• Filed: 03/14/2002
• Published: 03/13/2003
• Est. Priority Date: 09/12/1995
• Status: Active Grant

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Zhang, Lin, Velculescu, Victor E., Kinzler, Kenneth W., Vogelstein, Bert
• Granted Patent: US 6,746,845 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12N 15/1096 cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2539/103 Serial analysis of gene expression [SAGE]

Abstract

Serial analysis of gene expression, SAGE, a method for the rapid quantitative and qualitative analysis of transcripts is provided. Short defined sequence tags corresponding to expressed genes are isolated and analyzed. Sequencing of over 1,000 defined tags in a short period of time (e.g., hours) reveals a gene expression pattern characteristic of the function of a cell or tissue. Moreover, SAGE is useful as a gene discovery tool for the identification and isolation of novel sequence tags corresponding to novel transcripts and genes.

Claims

1. An isolated oligonucleotide composition derived from cDNA, said composition comprising:

at least two different defined nucleotide sequence tags, wherein each defined nucleotide sequence tag consists of about 6 to 30 nucleotides of said cDNA 5′

of the 5′

-most cleabage site of a restriction endonuclease within said cDNA or 3′

of the 3′

-most cleavage site of a restriction endonuclease within said cDNA;

wherein at least one tag corresponds to at least one expressed gene.