Mapping expressed sequence tag sites on yeast artificial chromosome clones of Arabidopsis thaliana DNA. | Zendy

Francis D. Agyare | Zendy; Deval Lashkari | Zendy; A Lagos | Zendy; Allen Namath | Zendy; Gus Lagos | Zendy; Ronald W. Davis | Zendy; Bertrand Lemieux | Zendy

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Mapping expressed sequence tag sites on yeast artificial chromosome clones of Arabidopsis thaliana DNA.

Author(s) -

Francis D. Agyare,

Deval Lashkari,

A Lagos,

Allen Namath,

Gus Lagos,

Ronald W. Davis,

Bertrand Lemieux

Publication year - 1997

Publication title -

genome research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.556

H-Index - 297

eISSN - 1549-5469

pISSN - 1088-9051

DOI - 10.1101/gr.7.1.1

Subject(s) - biology , yeast artificial chromosome , sequence tagged site , genetics , amplicon , genome , agarose gel electrophoresis , arabidopsis thaliana , expressed sequence tag , dna sequencing , gene , gene mapping , clone (java method) , chromosome , computational biology , microbiology and biotechnology , polymerase chain reaction , mutant

We describe a method for efficient parallel mapping of expressed sequence tag (EST) sites onto yeast artificial chromosome (YAC) clones. The strategy involves an initial YAC clone pooling scheme that minimizes the number of required PCR amplifications. This is followed by parallel analysis of PCR amplicons of EST sequences. Using this method, we have screened 600 EST sites in combinatorial pools of 3449 YAC clones that contain Arabidopsis thaliana DNA inserts. The presence of these genes on YACs was detected by amplifying EST sequences with PCR and analyzing the reaction products by agarose gel electrophoresis. Of the 600 ESTs, 271 were found to map to individual YACs. Software tools are presented that allow for the automated analysis of this electrophoresis data. Suggestions for the scale-up of this method to map large genomes are discussed.

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