BAC Clones Generated from Sheared DNA

Overview

Journal Genomics

Specialty Genetics

Date 2006 Nov 14

PMID 17098394

Citations 21

Authors

Kazutoyo Osoegawa

Gery M Vessere

Chung Li Shu

Roger A Hoskins

Jose P Abad

Beatriz de Pablos

Alfredo Villasante

Pieter J de Jong

Affiliations

Soon will be listed here.

Abstract

BAC libraries generated from restriction-digested genomic DNA display representational bias and lack some sequences. To facilitate completion of genome projects, procedures have been developed to create BACs from DNA physically sheared to create fragments extending up to 200 kb. The DNA fragments were repaired to create blunt ends and ligated to a new BAC vector. This approach has been tested by generating BAC libraries from Drosophila DNA with insert lengths between 50 and 150 kb. The libraries lack chimeric clone problems as determined by mapping paired BAC-end sequences to the assembled fly genome sequence. The utility of "sheared" libraries was demonstrated by closure of a previous clone gap and by isolation of clones from telomeric regions, which were notably absent from previous Drosophila BAC libraries.

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