Discovering distinct genes represented in 29,570 clones from infant brain cDNA libraries by applying sequencing by hybridization methodology.

  1. A Milosavljevic,
  2. M Zeremski,
  3. Z Strezoska,
  4. D Grujic,
  5. H Dyanov,
  6. S Batus,
  7. D Salbego,
  8. T Paunesku,
  9. M B Soares, and
  10. R Crkvenjakov
  1. Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, Illinois 60439, USA.

Abstract

To discover all distinct human genes and to determine their patterns of expression across different cell types, developmental stages, and physiological conditions, a procedure is needed for fast, mutual comparison of hundreds of thousands (and perhaps millions) of clones from cDNA libraries, as well as their comparison against data bases of sequenced DNA. In a pilot study, 29,570 clones in duplicate from both original and normalized, directional, infant brain cDNA libraries were hybridized with 107-215 heptamer oligonucleotide probes to obtain oligonucleotide sequence signatures (OSSs). The OSSs were compared and clustered based on mutual similarity into 16,741 clusters, each corresponding to a distinct cDNA. A number of distinct cDNAs were successfully recognized by matching their 107-probe OSSs against GenBank entries, indicating the possibility of sequence recognition with only a few hundred randomly chosen oligomers.

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