TOGA: an automated parsing technology for analyzing expression of nearly all genes.
We have developed an automated, high-throughput, systematic cDNA display method called TOGA, an acronym for total gene expression analysis. TOGA utilizes 8-nt sequences, comprised of a 4-nt restriction endonuclease cleavage site and adjacent 4-nt parsing sequences, and their distances from the 3' ends of mRNA molecules to give each mRNA species in an organism a single identity. The parsing sequences are used as parts of primer-binding sites in 256 PCR-based assays performed robotically on tissue extracts to determine simultaneously the presence and relative concentration of nearly every mRNA in the extracts, regardless of whether the mRNA has been discovered previously. Visualization of the electrophoretically separated fluorescent assay products from different extracts displayed via a Netscape browser-based graphical user interface allows the status of each mRNA to be compared among samples and its identity to be matched with sequences of known mRNAs compiled in databases.
Sutcliffe
J G
JG
Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. gregor@scripps.edu
Foye
P E
PE
Erlander
M G
MG
Hilbush
B S
BS
Bodzin
L J
LJ
Durham
J T
JT
Hasel
K W
KW
D001331:Automation; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics*; D020869:Gene Expression Profiling; D006801:Humans; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D012333:RNA, Messenger / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012984:Software; D014157:Transcription Factors / Q000235:genetics*
D004268:DNA-Binding Proteins; D016328:NF-kappa B; C539882:NR4A2 protein, human; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D012333:RNA, Messenger; D014157:Transcription Factors