High throughput direct end sequencing of BAC clones | Zendy

Jennifer Kelley | Zendy

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High throughput direct end sequencing of BAC clones

Author(s) -

Jennifer Kelley

Publication year - 1999

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/27.6.1539

Subject(s) - bacterial artificial chromosome , biology , deep sequencing , dna sequencing , genomic library , genetics , computational biology , contig , clone (java method) , insert (composites) , illumina dye sequencing , library , genome , gene , base sequence , engineering , mechanical engineering , 16s ribosomal rna

Libraries constructed in bacterial artificial chromosome (BAC) vectors have become the choice for clone sets in high throughput genomic sequencing projects primarily because of their high stability. BAC libraries have been proposed as a source for minimally over-lapping clones for sequencing large genomic regions, and the use of BAC end sequences (i.e. sequences adjoining the insert sites) has been proposed as a primary means for selecting minimally overlapping clones for sequencing large genomic regions. For this strategy to be effective, high throughput methods for BAC end sequencing of all the clones in deep coverage BAC libraries needed to be developed. Here we describe a low cost, efficient, 96 well procedure for BAC end sequencing. These methods allow us to generate BAC end sequences from human and Arabidoposis libraries with an average read length of >450 bases and with a single pass sequencing average accuracy of >98%. Application of BAC end sequences in genomic sequen-cing is discussed.

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