SBH and the integration of complementary approaches in the mapping, sequencing, and understanding of complex genomes
- Argonne National Lab., IL (United States)
A variant of sequencing by hybridization (SBH) is under development with a potential to inexpensively determine up to 100 million base pairs per year. The method comprises five experimental steps: (1) arraying short clones in 864-well plates; (2) growth of the M13 clones or PCR of the inserts; (3) automated spotting of DNAs by corresponding pin-arrays; (4) hybridization of dotted samples with 200--3,000 {sup 32}P or {sup 33}P-labeled 6- to 8-mer probes; and (5) scoring hybridization signals using storage phosphor plates. The method opens up intriguing possibilities for genome analysis. Some 200 7- to 8-mers can provide an inventory of the genes if cDNA clones are hybridized, or call define the order of 2-kb genomic clones, creating physical and structural maps with 100-bp resolution; the distribution of G+C, LINEs, SINEs, and gene families would be revealed. cDNAs that represent new genes and genomic clones in regions of interest selected by SBH can be sequenced by a gel method. Uniformly distributed clones from the previous step (20% of all) will be hybridized with 2,000--3,000 6 to 8-mers. As a result, approximately 50--60% of the genomic regions containing members of large repetitive and gene families and those families represented in GenBank would be completely sequenced.
- Research Organization:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- DOE Contract Number:
- W-31109-ENG-38
- OSTI ID:
- 37528
- Report Number(s):
- CONF-9206273-; ISBN 981-02-1157-0; TRN: IM9519%%452
- Resource Relation:
- Conference: 2. international conference on bioinformatics, supercomputing, and complex genome analysis, St. Petersburg, FL (United States), 4-7 Jun 1992; Other Information: PBD: 1993; Related Information: Is Part Of The second international conference on bioinformatics, supercomputing and complex genome analysis; Lim, H.A. [ed.] [Florida State Univ., Tallahassee, FL (United States). Supercomputer Computations Research Inst.]; Fickett, J.W. [ed.] [Los Alamos National Lab., Los Alamos, NM (United States). Center for Human Genome Studies]; Cantor, C.R. [ed.] [Boston Univ., MA (United States). Center for Advanced Research in Biotechnology]; Robbins, R.J. [ed.] [Johns Hopkins Univ., Baltimore, MD (United States). Applied Research Lab.]; PB: 672 p.
- Country of Publication:
- United States
- Language:
- English
Similar Records
SBH and the integration of complementary approaches in the mapping, sequencing, and understanding of complex genomes
Requirements in screening cDNA libraries for new genes and solutions offered by SBH technology