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Titolo:
Establishment of a high throughput EST sequencing system using poly(A) tail-removed, cDNA libraries and determination of 36 000 bovine ESTs
Autore:
Takasuga, A; Hirotsune, S; Itoh, R; Jitohzono, A; Suzuki, H; Aso, H; Sugimoto, Y;
Indirizzi:
Shirakawa Inst Anim Genet, Fukushima 9618061, Japan Shirakawa Inst Anim Genet Fukushima Japan 9618061 kushima 9618061, Japan Tohoku Univ, Fac Agr, Cellular Biol Lab, Aoba Ku, Sendai, Miyagi 9818555, Japan Tohoku Univ Sendai Miyagi Japan 9818555 Ku, Sendai, Miyagi 9818555, Japan
Titolo Testata:
NUCLEIC ACIDS RESEARCH
fascicolo: 22, volume: 29, anno: 2001,
pagine: NIL_8 - NIL_14
SICI:
0305-1048(20011115)29:22<NIL_8:EOAHTE>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
QUANTITATIVE TRAIT LOCI; MUSCLE MASS; IGF2 LOCUS; CATTLE; PIGS; MAPS; QTL; PROTEIN; GENOMES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
20
Recensione:
Indirizzi per estratti:
Indirizzo: Sugimoto, Y Shirakawa Inst Anim Genet, Fukushima 9618061, Japan Shirakawa Inst Anim Genet Fukushima Japan 9618061 061, Japan
Citazione:
A. Takasuga et al., "Establishment of a high throughput EST sequencing system using poly(A) tail-removed, cDNA libraries and determination of 36 000 bovine ESTs", NUCL ACID R, 29(22), 2001, pp. NIL_8-NIL_14

Abstract

We determined 36 310 bovine expressed sequence tag (EST) sequences using 10 different cDNA libraries. For massive EST sequencing, we devised a new system with two major features. First, we constructed cDNA libraries in whichthe poly(A) tails were removed using nested deletion at the 3'-ends. This permitted high quality reading of sequences from the X-end of the cDNA, which is otherwise difficult to do. Second, we Increased throughput by sequencing directly on templates generated by colony, PCR. Using this system, we determined 600 cDNA sequences per day. The read-out length was >450 bases in>90% of the sequences. Furthermore, we established a data management system for analyses, storage and manipulation of the sequence data. Finally, 16 358 non-redundant ESTs were derived from similar to 6900 independent genes. These data will facilitate, construction of a precise comparative map across mammalian species and isolate the functional genes that govern economic traits. This system is applicable to other organisms, including livestock, for which EST data are limited.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/05/20 alle ore 14:52:03